BISI NEW IT REPORT

TECHNICALREPORT ON STUDENTINDUSTRIALWORKEXPERIENCE SCHEME(SIWES) UNDERTAKENAT LAGOSSTATEUNIVERSITYHEALTHCENTER,OJO,LAGOS BY YUSUFAFEEZOLABISI MATRICNO:170561088 SUBMITTEDTO THEDEPARTMENTOFMICROBIOLOGY FACULTYOSSCIENCE LAGOSSTATEUNIVERSITY INPARTIALFULFILMENTOFTHEREQUIREMENTSFORTHE AWARDOFBACHELOROFSCIENCE(B.SC)DEGREEIN MICROBIOLOGY DECEMBER,2021

CERTIFICATION ThisistocertifythatthisreportoftheStudentIndustrialWorking ExperienceScheme(SIWES)programmewascarriedoutbyYUSUF AFEEZOLABISIStudentofLAGOSSTATEUNIVERSITY,OJO,LAGOS STATE,MatricNo.:170561088at,LAGOSSTATEUNIVERSITYHEALTH CENTER,Ojo,Lagos. _____________________________ _____________________________ Student’sSignature Supervisor’sSignature i

DEDICATION ThisreportisdedicatedtothealmightyGod,thegiverandsustainerof life,forHisunconditionalloveandmercygrantedtomethroughoutthe periodofmyIndustrialTraining. ii

ACKNOWLEDGEMENTS IgivethankstoAlmightyGod,whogavemethegiftoflife,and madeeverythingpossible. ThisacknowledgementwouldbeincompleteifIfailtoexpressmy gratitudetomylovelyfamilymostespeciallymyparents,MRANDMRS YUSUF,staffofthehealthclinic,tomentionbutafew,Mr.L.D OMOWAYEandMrs.A.IADEYEMIandMRS.ONWUTAasthechief medicallaboratorytechnologistandMRS.OBISESANasthechief medicalscientist,fortheirassistandsupportinthemedicallaboratory throughtheirrigoroustrainingcontributingtothesuccesstomy IndustrialTraining. iii

REPORTOVERVIEW TheStudentIndustrialWorkExperienceSchemeestablishedbythe FederalGovernmentofNigeriawasaimedatexposingstudentofhigher institutiontoacquireindustrialskillandpracticalexperienceintheir approvedcourseofstudyandalsotopreparestudentsfortheindustrial worksituationwhichtheyarelikelytomeetaftergraduation.This technicalreportisbasedontheexperiencesgainedduringmythree monthsofIndustrialTrainingatLagosStateUniversityHealthCenter, Lagos. Thisreporthighlightshowpatientsarebeingmanagedandalso theseveraltestcarriedoutforpatientssuchas:FullBloodCount(FBC), PackedCellVolume(PCV),WhiteBloodCellCount,DifferentialCount, StoolExamination,Microfilaria,Widal(Typhoidtest),Genotype,HIV,e.t.c. Iwasopportunedtoworkinfive(5)sectionswhicharePhlebotomy Section,HematologyandImmunohematology(BloodBank)Section, ClinicalchemistrySection,ClinicalMicrobiologySection,Serology Section.Thesesectionshaveexposedmetotheprecautions,rulesand regulationsofthelaboratory,howtodiagnosepatientsandhowthetests arebeinganalysed. Mostimportantly,itdescribestheactivitiesandmyexperience gainedduringtheperiodofthetraining,italsostatedtheproblems encounteredandalsogavesuggestionforimprovementofthescheme.

TABLEOFCONTENTS Titlepage Certification………………………………………………………….................i Dedication………………………………………………………………………ii Acknowledgements……………………………………………………………..iii ReportOverview…………………………………………………………..........iv CHAPTERONE 1.0 AbouttheIndustrialTrainingFund(I.T.F)……………………………1-3 1.1 AboutSIWES(StudentIndustrialTrainingFund)…………………1 1.2 Scope………………………………………………………………..1-2 1.3 Aim andObjectiveofS.I.W.E.S……………………………………2-3 1.4 HistoryandBackgroundofLagosStateuniversityhealthcentre…2-3 CHAPTERTWO 2.0 TheLaboratory…………………………………………………………...4-6 2.1 IntroductiontotheLaboratory……………………………………....4-5 2.2 SafetyRulesintheLaboratory………………………………………5 2.3 EmergencyintheLaboratory………………………………………..5 2.4 HazardousMaterials…………………………………………………5 2.5 HazardousEquipments…………………………………………...….5 2.6 LaboratoryEquipmentsandtheirUses………………………………5-6 CHAPTERTHREE 3.0 TheLaboratorySectionsandVariousTestsPerformed………………..7-28 3.1 ThePhlebotomyDepartment…………………………………………7-11 3.2 HematologyandImmunohematolgy(BloodBank)Section…………12- 16 3.3 SerologySection………………………………………………….….17-19 3.4 ClinicalchemistrySection…………………………………….…19-22 3.5 ClinicalMicrobiologySection………………………………….…….22-28 3.6 MicroscopyandCultures………………….…………………….……..28 CHAPTERFOUR 4.0 Summary,ChallengesEncountered,Recommendationand Conclusion………………………………………………………………..….29 4.1 Summary……………………………………………………….……...29 4.2 ChallengesEncountered………………………………………………29 4.3 Conclusion………………………………………………………….…29 4.4 Recommendation…………………………………………………...…29

CHAPTERONE 1.0 ABOUTTHEINDUSTRIALTRAININGFUND(I.T.F) InOctober1971,thefederalgovernmentestablishedtheIndustrialTraining Fund(I.T.F).Initspolicystatementno.1publishedin1973,aclausewasinserted dealingwiththeissueofpracticalskillsamongthelocallytrainedprofessionalsin thetertiaryinstitutionsespeciallytheUniversitiesofTechnology,Monotechnics, Polytechnics,CollegesofEducationandTechnicalColleges.Section15ofthe policystatementstatesclearlythat“Greatemphasiswillbeplacedonassisting certainproductsofthepost-secondaryschoolsystem toadaptororientateeasily totheirpossiblepostgraduationjobenvironments”,subsequentlyleadingtothe launchofaschemeknowastheStudent’sIndustrialworkExperience Scheme(SIWES). 1.1 ABOUTSIWES TheS.I.W.E.S.waslaunchedin1973bytheIndustrialTrainingFund(I.T.F)as aprogrammedesignedtoimparttheundergraduateofthenation’stertiary institutionsstudyingvariousprofessionalcourseswiththepracticalmethodsof performingprofessionalfunctionstoreallifesituationsonsite,intheofficeor eventhefactoryandhowtheyapplythemselvesmentally,intellectuallyand physicallyinrelationtowhattheyhavebeentaughtintheclassroomstheoretically. Itworkswiththefollowingprofessionalbodiestofunctioneffectivelyacrossthe tertiaryinstitutionsnationwide.ThesearetheNigeriaUniversityCommission (N.U.C),NationalBoardforTechnicalEducation(N.B.T.E.)andtheNational CommissionforCollegesofEducation(N.C.C.E.).Thus,equippingthestudents withthenecessaryskillsandtechnicalknowledgetomakethem highly competitiveandprofessionalindividualsintheLabourMarket 1.2 SCOPE TheschemeasconductedbytheIndustrialTrainingFund(I.T.F)throughtheir representativeliaisonunitsandofficessituatedwithinthevariousinstitutionand inmajorcitiesortownsinNigeriawiththenecessaryindustrialrudimentsneeded tocorroborate,practicalizeandthenactualizetherequiredtechnicalknowledge. TheIndustrialTrainingexperiencenotonlyputsthem inreallifesituationsbuts alsoexposestheirpracticalknowledgeofthecourseofstudy,consequently perfectingthisknowledgetherebyproducingverycompetentandversatile professionals. 1.3 AIM ANDOBJECTIVEOFS.I.W.E.S Theaim ofS.I.W.E.Sistobridgethegapbetweenthelevelofknowledge acquiredintertiaryinstitutionsandthepracticalapplicationofsuchknowledgein thefieldofwork. TheObjectivesare:  Toprovideanavenueforstudentsinindustriesofhigherlearningtoacquire industrialskillsandexperienceintheircourseofstudy.  Topreparestudentsfortheworksituationstheyaretomeetaftergraduation.

 Toexposestudentstoworkmethodsandtechniquesinhandlingequipment andmachinerythatmaynotavailableintheeducationalinstitution. Tomaketransitionfrom schooltotheworldofworkeasierandenhance studentscontactforlaterjobplacements.  Toimprovestudent’sinterpersonalrelationshipwithothersintheirfield.  Toprovestudentsanopportunitytoapplyhis/herknowledgeinrealworksituation, therebybridgingthegapbetweencollegeworkandactualpractice 1.4 HISTORYANDBACKGROUNDOFLAGOSSTATEUNIVERSITYHEALTHCENTER. TheHealthCentreisaprimaryhealthcarefacility,whichcatersformedical needsofstudents,staffmembersandtheirdependants,uptofour(4) membersthatarebelow18yearsold.TheHealthCentreconsistsofMedical, EnvironmentalHealth,Nursing,Pharmacy,X-ray,MedicalLaboratoryand MedicalRecordUnits.ItisheadedbyaMedicalDirectorwithabackgroundin PublicHealthwithassistanceofotherqualifieddoctorsandhealthworkers. Therearefacilitiestocaterforthestudents’populationandstaffonallthe campuses–Ojo,Epe,IkejaandJupeb,Badagry. VISION Tobetheforemostofpatientcentredcareprovider MISSION Toofferquality,holisticandexcellenthealthcareservicestoLASUcommunity andbeyond COREVALUE C–Compassion/Commitment A–Attention R–Respect E–Excellence ThecentreenjoysthesupportofLagosstateuniversityhealthcentersand LASUTHforreferralofcasesbeyonditsscope.Ithasfacilitiesforbothradio- diagnosisandmedicallaboratoryinvestigations.Itruns24hoursservicesand hasanAmbulancefortransferofcriticalcases.TheEnvironmentalHealth Unitofthecentreensuresallnuisancesintermsofrefuseandhazardous agents(livingandnonliving)oncampusareconstantlyabatedtoensure safety.Thereiscurrentlyacollaborativeeffortwithotherstakeholdersinthe healthsectorinLagosStateforanenhancedservicedelivery.

CHAPTERTWO 2.0 THELABORATORY 2.1 INTRODUCTIONTOTHELABORATORY Alaboratoryisafacilitythatprovidescontrolledconditionsinwhichscientific researches,experiments,andmeasurements,maybeperformed.Hencethe medicallaboratoryisalaboratorywheretestsarecarriedoutonclinical specimensinothertogetinformationaboutapatient’shealth. Therearefivesectionsinthelaboratory,theyare;PhlebotomySection, HematologyandImmunohematology(BloodBank)Section,Clinical chemistrySection,ClinicalMicrobiologySection,SerologySection.The overallsignificanceofthelaboratorydiagnosisisthattheyguidetowardsthe administrationofmosteffectivetherapysoastorestoreaproperhealthonthe patient.Laboratorysafetyprecautionsandethics 2.2 SAFETYRULESINTHELABORATORY Everylaboratoryisexpectedtoadoptacodeofbio-safetyprinciplesandwork practicewhichshouldbeenforcedandadheretostrictlybyworkersandvisitors. Allspecimenscomingintoandfrom thelaboratoryarebeingassumedtobe potentiallyinfectiousandharmfulandthatiswhythebelowprecautionsare ensuredtobetakentoavoidcontaminationandlaboratoryhazard.  AvoiddisruptinglaboratoryactivitiesyoumustTURNOFFallcellphonesand pagers:theiruseisprohibited.  Allpersonsinlaboratories,includingstudents,staff,andvisitors,shallwear safetyglasses,goggles,orfaceshieldsatalltimeswherepotentialeyehazards exist  Eating,drinking,chewinggum,andapplyingcosmeticsareprohibitedlaboratory.  Donotstorefoodorbeveragesinthesamerefrigeratorsorfreezerswith chemicals,biohazards,orradioactivematerials.  Neverconductunauthorizedexperimentsorengageinhorseplayinalaboratory. Pleaseimmediatelyreportanyunsafebehaviourtotheinstructor.  Wearappropriateclothing.Inparticular,youmustwearclosed-toedshoes(i.e., NOsandalsorflip-flops!)inthelaboratory.Ifyouhavealonghair,tieitback. Avoidwearingdanglingjewellery.  WearinganiPod,Bluetooth,oranyotherdevicethatinterfereswithhearingis notallowed.  Neverpipetteanythingbymouth.

 Theworkareamustbekeptcleananduncluttered.Allchemicalsshouldbe labelledandstoredproperly.  Thehazardsofchemicalsusedshouldbeknown(e.g.,corrosiveness, flammability,reactivity,stability,andtoxicity).  Alwayspayattentiontoyoursurroundingsandbeawareofwhatothersare doing.Alwaysbecourteous.  Removecontaminatedglovesbeforetouchingcommonusedevices(door knobs,faucets,equipment);discardglovesbeforeleavingthelaboratory.  Alwayswashhandsandarmswithantibacterialsoapandwaterbeforeleaving thelaboratory. Inconclusion,maintainingsafetyinthelaboratorylargelyrestonthe shoulderofthelaboratoryworkers.Adequatesafetyandgoodlaboratorypractice canbeavoidedirrespectiveofthelocation,staffstrengthandavailabilityof sophisticatedsafetycabinetsinthelaboratory.Whatarerequiredarehighly standardsofhygienebythelaboratoryworkerstoachievegoodresultsintheir dailyoccupationalpractice. 2.3 EMERGENCYINTHELABORATORY  Knowwheretofindthenearestexitincaseoffireorotheremergency.  Knowthewhereaboutsofthenearestfireextinguisher,fireblanket,firstaidkit, eyewashequipment,showerandtelephone.  Incaseoffire,clearoutofthelaboratoryfirst,andthencallanemergency number. 2.4 HAZARDOUSMATERIALS  Bothliquidanddrychemicalscanbeflammable,poisonous,carcinogenic,etc. Payattentiontospecialinstructions,suchasto;workwithasubstanceonlyin afumehood.  Biologicalhazardsincludebacteriaandbodyfluids,suchasblood.Handle withappropriatecare,anddisposeofbiologicalhazardsasinstructed.  Disposeofhazardousmaterialsasinstructed.Neverputanythingdownthe sinkwithoutcheckingwithaninstructor.  Cleanupspillsandbrokenglass.Don'thandlebrokenglasswithyourbare hands.Useabroom anddustpan,andthrowawayallbrokenglassand disposableglasspipettes,coverslips,andothersharporeasilybreakable glassinacontainerforglassdisposalonly.Notifytheinstructorimmediately ofallincidents. 2.5 HAZARDOUSEQUIPMENTS  Ifappropriate,turnoffequipmentthatisn'tbeingused.  DonotuseaBunsenburnerunlessinstructedtodoso.  Keepliquidsandchemicals,especiallyflammablematerials,wellawayfrom

anyheatsourceorelectricalequipment.  Ifanyelectricalequipmentismalfunctioning,makingstrangenoises,sparking, smoking,orsmells\"funny,\"donotattempttoshutitofforunplugit.Getan instructorimmediately.Itisimperativethattheinstructorknowofany equipmentproblems. 2.6 LABORATORYEQUIPMENTSANDTHEIRUSES  Microscope:Isusedtoexaminesamplesandtoanalyzetheircontentsthatare notvisibletothenakedeye.Itisalsousedtocountpathogenandothercells andtoviewunderx10,x40,andx100objectives.  Autoclave:ForSterilization.  Centrifuge:Isusedforspinningspecimene.g.urinetoenableseparationinto constituentsorcomponentse.g.bloodintoserum andplasma.  Refrigerator:Providessuitabletemperatureforstorageandpreservationof reagents,unusedmedia,bloodsamplesetc.  Bunsenburner:Servesasthesourceofheatforsterilizingwireloop,surgical forcepsandothermetalinstrumentstobeusedforanalysis.  WeighingBalance:Usedformeasurement.  Wireloop:Itisusedforstreakingspecimenoncultureplatesanditcanalsobe usedformakingsmearofsamplesonslides.  Lancet:Itisasterileneedleusedtoprickthethumbforthecollectionofblood samples.  Capillarytube:Itisusedforthecollectionofbloodsamplestodeterminethe packedcellvolume.  Universalbottle:Usedforsamplecollectione.g.urine,stool,semen  Glassslide:Itisusedforthepreparationofsamplestobevieweddirectlyunder themicroscope. Sterileswabstick:Isusedforthecollectionofsamplestodirectlyfrom the sightofinfectione.g.Ear,nose,vagina,cervix,etc.  Samplingbottles:Theyarebottlesusedforthecollectionofbloodsamplese.g. universalbottle,fluorideoxalatebottle,Ethylene-Di-amine-TetraaceticAcid bottle(EDTA),Lithium Heparinbottle,plainbottle.  Incubator:Usedforculturingordryingofmicroorganism.  Microheamatocritcentrifugemachine:Itisusedtospinsampleforthe analysisofpackedcellvolumeofbloodsample.

 Waterbath:Useasheatingapparatus  Microhaematocritreader:Usedtoreadthepackedcellvolumeinpercentage.  Tourniquet:Itistightenedonpatienthandinthecollectionofbloodsamplein ordertogetaprominentveinbeforeincision.  NeedleandSyringe:Itisusedforthecollectionofbloodsamples.  Macrocentrifugemachine:Itisusedfortheseparationofbloodsamplesin ordertogettheplasmaandalsousedfortheseparationofurinesamplesoas togetthesupernatantandthespecimen  Glucometer:Usedtocheckforthesugarlevelinthebodywiththeaidofits strip.  Hematologyanalyzer:ItisusedfortheanalysisofFullBloodCount(FBC).

CHAPTERTHREE 3.0 THELABORATORYSECTIONSANDVARIOUSTESTSPERFORMED 3.1 THEPHLEBOTOMYDEPARTMENT ThePhlebotomyprocedurefacilitatesobtaininggoodqualityspecimenson thecorrectpatientforfurtheranalysisinthelaboratory 3.1.1 IDENTIFICATIONOFMATERIALSUSEDINTHEPHLEBOTOMYANDUSES 1.PersonalProtectiveEquipment  PhlebotomyUniform:Serveasprotectivetothebody  Disposablegloves/HandGel:Forprotectionagainstspillage 2.Needles:Usedtomakeincision 3.Needleholders:Forholdingneedleoralsoknownasvenoject 4.Vacutainers,VacutainerholderorSyringe:Servesasblooddrawer 5.SamplebottlesaccordingtoOrderofDraw:  EDTA(EthyleneDiamineTetraAceticAcid) Alkalinephosphatase-duetochelationofmetalliccofactorswhichinhibits theenzymeactivity.Potassium andsodium -duetotheadditionof potassium tothesample.Calcium andmagnesium -thesearechelatedby theEDTA. EDTAisalsoanunsuitableadditiveforsamplesrequiringbacterialculture, sincethechelationofthedivalentcationsinhibitsthegrowthofbacteria. EDTAissometimesusedtopreventcellsclumpinginfluidsamples requiringcellcountstoaccompanyacytologyevaluationbutitdoesnot actually'fix'thecells.Asamplefixedinformalinoralcohol/ethanolis requiredforaccuratecytologyexamination.EDTAisnotsuitablefor samplesrequiringvirusisolationincell/tissueculturebecauseitformsa gelwhenaddedtothecellculturemedium andthisdisruptsthecell monolayer.  Plain(noanticoagulant) Plain(orclotted)samplesareusedtoprovideserum forserologyand mostbiochemistryorendocrineassays.Serum isplasmawithout fibrinogensincethefibrinogenhasbeenused'intheformationoftheclot.  Lithium Heparin Lithium heparinacceleratestheactionofantithrombinIIIwhichneutralizes thrombinandthuspreventstheformationoffibrinfrom fibrinogen(clot formation).Thiseffectmakesheparinsamplesunsuitablefor determinationoffibrinogenorclottingfactors. Lithium heparinisastandardanticoagulantusedtoobtainplasmafor biochemistryanalysis.Lithium heparinisthemostsuitableanticoagulant fortheisolationofvirusesincell/tissueculture.Thisanticoagulantisnot suitableforhaematologyastheheparinaltersthecellmorphology.Whilst measurementofhaemoglobinandbloodcellcountscanbeobtainedusing

thisanticoagulantanaccuratewhitecelldifferentialandmorphology commentsarenotpossible.  Fluoride/Oxalate Fluoride/oxalatesamplesareusedforglucose(andlactate)determination only.Sodium fluoridefunctionsbystabilizingthebloodcellmembraneand inhibitingtheenzymesystemsinvolvedinglycolysis,whichpreventsred bloodcellsmetabolisinganyglucosepresentinthesample.Forthis reasonitistheonlysuitablesampleforaccurateglucoseanalysis. Fluorideisapotentinhibitorofmanyenzymesandtheinhibitionof glycolysistendstocausefluidshifts.Fluorideisaweakanticoagulanton itsown,sopotassium oxalate(anotherpowerfulenzymeinhibitor)is usuallyaddedtosupplementitsaction.Otherplasmaorserum samples maybeusedforglucoseanalysisONLYiftheplasma/serum isseparated from thecellswithin1hourofsamplecollection.Withoutanantiglycolytic agent,thebloodglucoseconcentrationdecreasesapproximatelyby0.56 mmol/lperhourat25°C.  Sodium citrate Sodium citrateisthestandardanticoagulantforcoagulationassays. Citratefunctionsbychelatingcalcium.Theeffectiseasilyreversedbythe additionofcalcium tothesample.Otheranticoagulantsarenotsuitable forcoagulationassaysastheyinterferewithcoagulationfactors.Citrate alsoinhibitsALP,ALTandinterfereswiththemeasurementofphosphate.

6.Tourniquet:Usedtosearchvein.Prolongedvenousocclusioncancause changesinconcentrationsofbloodconstituents.Therefore,theuseofa tourniquetshouldbeminimized.Ifatourniquetisusedtosearchforavein,it shouldbereleasedbeforewithdrawalofbloodbegins.Inanycase,theuseof atourniquetshouldbelimitedtolessthanoneminute. 7.70%Alcoholswabs:Usedasadisinfectantfortheareainwhichincisionisto bemade. 8.Microporetape:Toaidhemeostatis 9.Dentalrolls 10.Adhesivedressing 11.Racks 12.Disposablewastebins 13.CottonwoolPilloworothersupport:Usedforsupportingthearm foreasy bloodflow 3.1.2 STANDARDOPERATIONPROCEDURE(S.O.P)FORBLOODCOLLECTION Thefrequentpointofbloodcollectionisusuallyfrom thevein(venipuncture). Thematerialsforthepatient’sidentitymustbecheckedbeforeattempting venepuncture.ThismustbecarriedoutbyaskingthepatienttheirFullName andDateofBirth.  CheckthatthisinformationcorrespondswiththatontheRequestform. AnyamendmenttothesedetailsoranyothersontheRequestform mustbein accordancewiththeDirectoratePolicy.  Wherepatientdetailslacklegibility,staffmaywritethecorrectdetailsclearly nexttothoseontheform withoutcrossingouttheoriginaldetails.  Iftestsarerequestedthatareunfamiliarandstaffareunsureoftheappropriate bloodtubesforspecimenscheckthelist‘Whattubeguide’availableateach workstationor,whennecessarycontactaqualifiedBiomedicalScientistinthe appropriatedepartmentwithinthePathologyLaboratory.  Examinebotharmsofthepatientandselecttheonethatappearsappropriate Ensurethatthepatientiscomfortableandthatthearm iswellsupportedand examinepotentialvenepuncture. Askthepatienttobareanarm,ensurethatthearm iswellsupportedand applythetourniquettothepatient’sarm,justabovetheelbowandtight enoughtoallowtwofingersbehindthestrap.

 Tightenthetourniquetalittlemore,takingcarenottopinchtheskin Askthepatienttostraightentheirarm andclenchtheirfist.Thiswillmake theveinmoreprominent.  Ifnecessaryrubthebendoftheelbowtomaketheveinmorevisible.  Feelwithafingertipforthe‘best’veinatthebendoftheelbowratherthan plungetheneedleintoapoorveinthatlooks‘alright’. Ifthisfailsasuitableveincanoftenbefoundatthesideofthearm onthe elbowside. Itmaybenecessary,onoccasiontotakebloodfrom thebackofhand.  Applythetourniquetabovetheelbow.Thetourniquetisclosedaroundthearm byinsertingtheplasticclipintotheholderandthentightenedappropriatelyby pullingthestrap. Askthepatienttostraightentheirarm andtomakeafistinordertomakethe veinsmoreprominent.  Feelwiththefingertipifnecessarytolocateasuitableveintopuncture.  Ensurethatequipmentandbloodtubesrequiredareimmediatelywithineasy reach.  RemovethetopplasticsectionofaVacutainermulti-sampleneedleandscrew threadintoaVacutainerneedleholder.  Disinfectsitewitha70%Isopropylalcoholswab.  Leavefor30secondsforthealcoholtoevaporateandduringthisperiod assemblethebloodtubesrequired. Removethecoverfrom themulti-sampleneedleanddiscardintoaclinical wastebin.  Keepingtheneedleholderandattachedmulti-sampleneedleinonehanduse thethumbontheotherhandtopressontheveinjustabovethechosenentry pointandpulltheskinbackslightlytowardsyoutoholdtheveinfirmlyand stretchtheskinoverthechosensite.  Withtheneedleholderandmulti-sampleneedlealmostparalleltothepatient’s arm andtheneedlebeveluppermost,gentlypushtheneedleintothechosen venepuncturesite.  Onceintheveinholdtheneedle-holdersteadyandgentlypushthecapofthe appropriatebloodsampletubeontothecoveredsampleneedleatthebaseof theinsideoftheneedle-holder.  Bloodshouldenterthesampletubeandfilltotheappropriatelevelindicated.

Removethesampletubefrom thesampleneedlewhenfullandattachanother sampletubeinrequired.  Bloodsamplesmustbegentlymixedattheearliestopportunitytoensure anticoagulationeffectiveness  Asthelastbloodsampletubeisfillingslackenthetourniquetbypressingdown onthereleaseclipthatisonthesideawayfrom thearm. Withdrawtheneedlefrom theveinandquicklyapplyacleanpadofcottonwool.  Askthepatienttokeeppressureonthecottonwooltostopfurtherbleeding.  DiscardtheneedleandholderintoaSharpsbin.  Gentlymixthesampletube(s).  Ifthepatientisunabletomaintainsufficientpressureonthevene-puncturesite applythispressureforthem. Removethetourniquetfrom thepatient’sarm. Whenbleedingfrom thevenepuncturesitehasstoppedapplyMicroporetape tightlyoverthecottonwool.  Theprocedureisnowcompleteandthepatientcanleave.

3.1.3PHLEBOTOMYDIAGRAMS

3.2 HEMATOLOGYANDIMMUNOHEMATOLGY(BLOODBANK)SECTION Inhematologysection,theanalysisiscarriedoutusingthewholeblood sampleofpatientfordiagnosisofhematologicaldiseasesandabnormalities. BloodsamplesarecollectedinEDTAbottleforanalysis. ImmunohematologySectionAlsoknownasthebloodbankperformsteststo providebloodandbloodproductstopatientsfortransfusionpurposes.Theblood banktechnologistreliesonthephlebotomisttoperform identificationofthe patientwithouterror,sincepatientswilldieifgiventhewrongbloodtype.The analysescarriedoutinthesesectionsinclude:Packedcellvolume,Fullblood count,Erythrocytesedimentationrate(ESR),Bloodfilm formicrofilariaand ABO/D(Rh)typing,Antigentyping,Bloodgrouping,Crossmatching,respectively. 3.2.1 MATERIALSUSEDINHAEMATOLOGYANDIMMUNOHEMATOLOGY SECTION Pipette,HaematocritcentrifugemachineforPCV,Haematocritcentrifugereader forPCV,MicroHaematocritanalyzerforFullBloodCount,Macro,Microscope, Microscopyslide,Electrophoresismachine,Coverslip,Bunsenburner,Plasticine, Sterilecapillarytube,Washbottle,Westergrentubes,Stopwatch,Testtubes, Refrigerator,Racks,Variousdisposablewastebins,Tiles,Scissors,Ethylene diaminetetraaceticacid(EDTA),Leishmanstain,Normalsaline,Water,Antisera forbloodgroup,Buffersolution,Oilimmersion. 3.2.2 COMPLETEBLOODCOUNT(CBC)TEST  Introduction:Thecompletebloodcountofabloodsamplehelpstoknowthe totalcellinthewholeblood.Itdeterminesthetotalhaematocrit(HCT), hemoglobin(HGB),redbloodcell(RBC)count,whitebloodcell(WBC)count, plateletcount,meancorpuscularhemoglobin(MCH),meancorpuscular hemoglobinconcentration(MCHC),meancorpuscularvolume(MCV), differential(DIFF)-doneonabloodsmear  Aim:Todeducethetotalcountsofallbloodcomponents  Equipments/Apparatus:Hematologyanalyzer,wholebloodsampleinanEDTA bottle.

 Procedure:BloodsamplewascollectedintoanEDTAbottle(Lavender StopperedTube)throughvenipunctureandwasmixedwithanticoagulantby invertingthebottlegently8times.Thebloodsamplewasplacedunderthe hematologyanalyzersensitiveprobe.Theprobebuttonwaspressedsothatthe probecanpickthesampleintothemachineforanalysis.Theresultwas displayedonthescreenofthemachineandthenprintedout.  Conclusion:Thecountofplatelet,whitebloodcellanddifferentials, haemoglobin,granulocyteandallothercellsinthebloodsampleswas determined 3.2.3 PACKEDCELLVOLUME(PCV)TEST  Introduction:Thepackedcellvolumeisthevolumeoccupiedbythepackedred cellafteravolumeofanti-coagulatedvenousbloodisfullycentrifugedinto plasmaandredbloodcell.Thevolumeofpackedcellisexpressedasa percentageoftheoriginalvolumeoftheblood.  Aim:Toestimatetherelativemassofredbloodcellspresentinabloodsample inpercentagevolume.  Equipments/Materials:Haematocritreader,Bunsenburner,Microhaematocrit centrifuge,Heparinisedcapillarytube,wholebloodinananticoagulantbottle (EDTA),Microhaematocritreader,anabsorbentcottonwool.

 Procedures:ThebloodsamplewascollectedintoanEDTAbottle.The heparinizedcapillarytubewasfilledto2/3lengthofthetubefrom theblood sampleandOneendofthetubewassealedwithflameusingtheBunsenburner, thenabsorbentcottonwoolwasusedincleaningthetubebeforeplacinginthe centrifuge.Thesealedtubewasplacedinthemicro-haematocritcentrifuge machine,therebyplacingthesealedendoutwardtotouchthebaseofthe spinner.Thesealedtubewasspuninthehaematocritcentrifugeat 12,000/13,000rpm for5minutes.Thespuntubewasplacedonthemicro haematocritreadertoreadtheresultinpercentage,positionedinslotsothat thebaselineintersectsthebaseofredcellsandtubeholderwasmovedsothat thetoplineintersectsthetopofplasma,thenknobwasadjustedsothatthe middlelineintersectsthetopofredcell. Thepercentagepackedcellvolumeonthescalewasread.  Result: Adult: Normalrangeformale 37-50% Normalrangeforfemale 35-45% Children: NormalRange 29-41%  Conclusion: FactorsaffectingtheaccuracyofPCVare;Unsteadypowersupply,Poorblood samplecollection,Parallaxerrorwhilereadingtheresultonthehaematocrit reader,Incorrectbloodtoanticoagulantratio,Overspinningofthebloodinthe centrifuge,Lysingofthebloodbyflameordelayinspinning Bio-medicalsignificance:LowPCVvalueindicateshortageofblood 3.2.4 ERYTHROCYTESEDIMENTATIONRATE(ESR)  Introduction:ErythrocyteSedimentationratetestisusedtodetect inflammationinfections,cancer,andautoimmunediseases;itisalsouseto monitorwhetheranillnessisrespondingtotreatment.Itisascreeningtestso cannotbeusedtodiagnoseaspecificdisorder  Aim:TodeterminetherateofinflammationofbloodusingWestergrenmethod. Equipment/Apparatus:Sodium Citratebottle,ESRrack,Westergrenglassrod.

 Procedure:BloodsamplewascollectedintoanEDTAbottleandinverted8 timestoenabletheactionoftheadditiveandthenpouredintotheWestergren tubecontainingsodium citrateandwasmixedgentlyagainbyinvertingthetube 8time.TheWestergrenglassrodwasforcedintothetubeandbloodrisesto zeromark.ThesamplewasplacedinaverticalstandontheESRrackandtimed forone-hour.Theresultwasrecordedjustafterone-hourinmillimeters  Result:Thenormalrangeofthesedimentationinadult Menunder50years………………………..lessthan15mm/hr Menover50years………………………….lessthan20mm/hr Womenunder50years………………..........lessthan20mm/hr Womenover50years……………………….lessthan30mm/hr Thenormalrangeofthesedimentationinchildren Newborn…………………………………………lessthan2mm/hr Neonataltopuberty……………………………..3-13mm/hr  Conclusion:TheESRtestdoesnotdiagnoseaparticularkindofinfectionbut rathertelliffurthermedicationshouldbegiven.Itisusuallydonetoindicateif thereareabnormalproteinsinthebody 3.2.5 BLOODGROUPINGANDGENOTYPINGTEST Introduction:BloodgroupingoftheABOsystem isdeterminedwithAnti-A,Anti -B,andAnti-Dsera,whichform agglutinationcomplexwithantibodiesfoundin thebloodsample.  Aim:Todeterminethegroupandtherhesusofapatient’sblood  Equipments/Materials:Cleanfreegreasetile,Pasteurpipette,Wholeblood sampleinanEDTAbottle,distilledwater,applicatorstick,testtuberack, electrophoresismachineandtank,cleanwhitetile,cottonwool,applicatorstick, cellulosefilterpaper,gloves. Reagents: Anti-A,Anti-B,Anti-Dsera,Bufferforbalancing,normalsaline

 Procedure: ForbloodGrouping:ThebloodsamplewascollectedintoanEDTAbottle throughvenipuncture.10ulofbloodwasplaced3spotsonthetilewiththeaid ofPasteurpipette.TheantiseraA,BandDwereplacedcarefullyoneachspots, ABOofthegroupingsystem onthetilerespectivelyandanapplicatorstickwas usedtothoroughlymixthedropofbloodwiththeanti-seraoneaftertheother withoutcontamination.Thetilewasgentlyrockedfrom sidetosidefor3 minutestoallowagglutinationoccurrence,thenresultwasobserved. ForGenotyping:Cellswerewashedtwotothreetimesinatesttubecontaining normalsalineafterwhich,adropofwashedcellswereplacedonatile.Thisis followedbythehemolysisofbloodonthetileandtheplacementofASandAA controlusingapplicatorstick,aftermakingsurethatthebufferinsidethe electrophoresistankcoveredtheelectrode,thecellulosepaperwasplacedon thetank,whichisthencoveredandmains(current)switchedon.Readingwas recordedafter5-10mins  Result:Theresultforbloodgenotypewastakenbystudyingthemovementand separationofhemoglobinmolecule.

 Conclusion: Theresultwasobservedaccordingtotheagglutinationthat occurredineachspotsonthetile.AntiDdeterminesthepresentoftherhesus ‘D’factorinbloodgroup. Factorsthataffectbloodgroupingare;wronglabelingofspotandconfusionof anti-serawithspots,Contaminationoftestcardortileswithdetergents,Expired anti-sera Bio-medicalsignificance;Bloodtransfusion,Bloodcompatibility,Antenatal screening# 3.3 SEROLOGYSECTION Testsdoneinthisdepartmentaredesignedtodetectthebody'sresponsetothe presenceofbacterial,viral,fungal,parasiticandotherconditionswhichstimulate detectableantigen-antibodyreactionsinatestsystem toaidinthediagnosisof thepatient.Mosttestsperformedinthissectionarecarriedoutunderthe principlesofImmunoassay,someofthem are;Coldagglutinins(CAG)-specimen mustbekeptwarm,Rheumatoidarthritis(RA),VDRL,todiagnosesyphilis, PregnancyTesting,Widal 3.3.1 HBs.AgTESTFORHEPATITIS,VDRL(VENERALDISEASESRESEARCH LABORATORY)TESTFORSYPHILISUSINGSTRIPS  Introduction:HBsAGisarapidimmunochromatographictestforthequalitative detectionofHepatitisBsurfaceAntigeninhumanserum/plasma,itcanbe usedforprenatalortransfusionscreening,andduringacuteinfectionorchronic carriageoftheHepatitisBvirus.Earlydetectionofinfectionisessentialfor rapidinitiationofadequatetreatment. VDRLtestisascreeningtestforsyphilis.Itmeasuressubstancescalled antibodiesthatbodymayproduceifitcomesincontactwiththecausative agentofsyphilis,whichiscalledTreponemapallidium Aim: Todeterminethepresenceorabsenceofhepatitisandsyphilisinthe bodysystem. Materials: HBsAGTeststrips,VDRLteststripEDTAbottle,Centrifuge,clean testtube Specimen: Serum. Procedure: Thepatientbloodsamplewascollectedintoaplainbottlethrough venipuncture.Thebloodsamplewasspuninacentrifugefor5minutes,after spinningtheserum wasseparatedcarefullyintoacleantesttubebytheuseof Pasteurpipetteandthenteststripwasimmersedverticallyintotheserum for 10minutes.Theobservationwastakenafter10mins.  Result:AppearanceofalineattheControlregionandanotherattheTest indicatespositiveresult,whileanappearanceofalineattheControlregiononly, indicatesnegativeresult.Whenthereisnoappearanceofanyline,meansthe

testininvalidandastoberedoneusingnewkits. 3.3.2 BLOODPREGNANCYANDURINEPREGNANCYTEST,USINGTESTSTRIP.  Introduction:Apregnancytestisdonetodetermineifawomanispregnant, pregnancyhormonecalledtheHumanChorionicGonadotrophin(HCG)intothe bloodandurine.PregnancytestdetectsthehormoneHCGandconfirmsthe pregnancy.  Aim:todeterminethepresenceofpregnancyhormone(HCG)inthebloodand urine.  Materials:pregnancyteststrip,plainbottle,needleandsyringe,wetswab, cottonwool,centrifuge,cleantesttube.  Specimen:blood(serum)andurine  ProcedureforBloodPregnancyTest:Patient’sbloodwascollectedthrough venipunctureintoplainbottle,bloodsamplewasspuninacentrifugefor5 minutes,andtheserum wasseparatedcarefullyintoacleantesttubebythe useofPasteurpipette.Thepregnancyteststripwasimmersedverticallyinto theserum for5minutes.Thestripwasremovedandthereactionwasobserved.  ProcedureforUrinePregnancyTest:Thepatient’surinesamplewascollected intouniversalsterilebottleandthepregnancyteststripsawimmersedintothe urinefor3seconds,thenremovedandleftfor5minsandtheresultwas observed.  Result:AnappearanceofalineattheControlregionandanotherattheTest indicatespositiveresult,whileanappearanceofalineattheControlregiononly, indicatesnegativeresult.Whenthereisnoappearanceofanyline,meansthe testininvalidandastoberedoneusingnewkits

3.3.3 WIDALTEST  Introduction:TyphoidfeverisaninfectiousdiseasecausedbytheSalmonella typhi,itisdiagnosedbyWidaltestwhichemploysanantigen-antibodyreaction toscreenforthepresenceofSalmonellatyphiandparatyphiantibodiesinthe sampleserum..theorganism istransmittedbywaterorfoodcontaminatedby faecesoftyphoidvictimsorcarriers,thatisapersonwhoharboritwithout showingsignsandsymptoms.  Aim:ToinvestigatethepresenceofSalmonellatyphiandparatyphiintheserum ofpatient.  Materials:Testcard/whitetiles,Pasteurpipette,centrifuge,antigenkitandstop watch. Procedure:3-5mlofbloodwascollectedfrom thepatientthroughvenipuncture intoaplainbottleandthebloodwasspunat3000revperminfor5minutesso astoseparateouttheplasma.Adropperwasusedtocarefullydrawnthe antigenkitsandadropwasplacedoneachofthetestcardinpairsoffour spotslabeledO,OA,OB,OCandH,HA,HB,HCandadropofserum was carefullyaddedintotheantigenrespectivelywiththeaidofPasteurpipetteand mixedtogetherwiththeaidofanapplicatorstickthetestcardwasrocked gently,therateofreactionandagglutinationwasobservedatanintervalof 30sec,1min,2mins,and5mins  Result Reactive:visibleagglutinationonspotHandothersindicatethepresentof Salmonellaantibodies Nonreactive:novisibleagglutinationindicatesabsenceofSalmonella antibodies

Widaltest: Positive Highlyreactive……………………………….1:320(agglutinationreactionbefore60 seconds) Veryreactive……………………………….1:160(agglutinationreactionbefore120 seconds) Reactive………………………………………1:80(agglutinationreactionbefore180seconds) Widaltest:Negative Nonsignificant…………………………………1:40 Nonsignificant………………………………….1:20 Notreactive…………………………………….Nil 3.3.4RETROVIRALTEST  Introduction:ThisisthediagnosisforHumanImmunodeficiencyVirus,an infectiousagentthatcausesAcquiredImmunodeficiencySyndrome(AIDS),a diseasethatleavesapersonvulnerabletolifethreateninginfection.HIV transmissionoccurswhenapersonid=sexposedtobodyfluidsinfectedwith virus,suchasblood,semen,vaginalsecretionsandbreastmilk. Aim: ToinvestigatethepresenceofHIV1and2inpatient’sblood  Materials:Determinekits,Unigoldkit,StatpackBuffer,Plainbottle,pipette Specimenused: Serum  Procedures:Thebloodsample,collectedinaplainbottlewascentrifugedat 3000rev/mintoallowseparation.Theserum waspickedwithapipetteandtwo dropswasplacedonthesamplepadofthedeterminekitandallowedto penetratethenleftfor15min.Ifresultprovespositive,theUnigoldkitswouldbe usedfollowingthesameprocedure.AfterusingUnigoldtoconfirm theresult andprovesnegativetheStatpackitwouldbeusedtoasaconfirmer.  Result:TheappearanceofalineattheControlregionandanotherattheTest regionindicatesaPositiveresult,whiletheappearanceofalineontheControl regiononly,andindicatesaNegativeresult. 3.4 CLINICALBIOCHEMISTRYSECTION Thissectiondealswithchemicalanalysisofserum orplasmainwhichmany diseasesofthemajororganssystemscanbediagnosedsuchasheartattacks, hepatitis,renalfailure,diabetes,Liverfunction,etcBloodsamplesamplesmay collectedintotheSerum SeparatorTubeorLithium Heparin.Testperformedin thisdepartmentare:  BloodGlucose;FBS,FBS2HPP,RBS,OGTT  Bloodlipids(fat)Cholesterollevel.  Electrolytes-sodium,potassium,CO2-(Bicarbonate),andchloride

 Uricacid  CreatinineandBloodUreaNitrogen(BUN)  Liverfunctiontests-AST,ALT,alkalinephosphatase,andbilirubin. 3.4.1 MATERIALSANDEQUIPMENTSUSEDINCLINICALBIOCHEMISTRYSECTION PersonalProtectiveEquipments(PPE),BloodcollectionmaterialsDifferenttubes like;Lithium Heparin,Serum SeparatorandFluorideOxalatetubes,Chemical reagentsanddetergents,automatedmachines,centrifuge,GlucometerandAccu check. 3.4.2 BLOODGLUCOSE TestOverview:Abloodglucosetestmeasurestheamountofatypeofsugar, calledglucose,inyourblood.Glucosecomesfrom carbohydratefoods.Itisthe mainsourceofenergyusedbythebody.Insulinisahormonethathelpsyourbody cellsusestheglucose.Insulinisproducedinthepancreasandreleasedintothe bloodwhentheamountofglucoseinthebloodrises. Normally,yourbloodglucoselevelsincreaseslightlyafteryoueat.Thisincrease causesyourpancreastoreleaseinsulinsothatyourbloodglucoselevelsdonot gettoohigh.Bloodglucoselevelsthatremainhighovertimecandamageyour eyes,kidneys,nerves,andbloodvessels. Thereareseveraldifferenttypesofbloodglucosetests. 1.Fastingbloodsugar(FBS)measuresbloodglucoseafteryouhavenoteatenforat least8hoursandatmost14hours.Itisoftenthefirsttestdonetocheckfor prediabetesanddiabetesMaterials/Reagents:FluorideOxalateandotherblood collectionequipments,Centrifuge,Insulinkit(NORUDIA® Insulin)Liquidreagent, automatedmachine,GlucometerorAccuCheck. Procedure:Patientsisensuredtohavefastedfortherequiredperiodoftime beforesamplecollectionintoaFluorideOxalatetube,andthenthebloodsamples arecentrifugedinordertoobtainplasma.Theplasmaobtainedwasdecantedinto asmallcapwhichisthentransportedintothemachineforanalysis.Theresults arethendeducedwhichismeasuredinmg/dl NormalResult:Normalresultislessthanorequalto100mg/dl 2.2-hourspost-prandialbloodsugarmeasuresbloodglucoseexactly2hoursafter youstarteatingameal.Thisisnotatestusedtodiagnosediabetes. Procedure:AfterperformingthesameprocedureforFBSsamplecollection, patientsarethenaskedtoreturn2hrsassoonastheystarteating,thenanother venepuctureismadeandthesameprocedureisrepeated. Result:Normalresultsislessthan140mg/dlforpeopleage50andyounger;less than150mg/dlforage50-60;lessthan160mg/dlforage60andolder. 3.Random bloodsugar(RBS)alsoknownascasualbloodglucosetest.RBS measures Bloodglucoseregardlessofwhenyoulastate.Severalrandom measurements

maybetakenthroughouttheday.Random testingisusefulbecauseglucose levelsinhealthypeopledonotvarywidelythroughouttheday.Bloodglucose levelsthatvarywidelymaymeanaproblem. Materials/Reagents:SamematerialsasFBS. Procedure:GlucometerorAccuCheckaremostlyusedtoperform thistest.The Glucometercorrespondingcodestripisinsertedandloaded,Patient’sthumbis disinfectedusinga70%alcoholpadandprickedwithlancet.Thebloodiswipedoff inordertoavoidsamplingerror,pressureisappliedbelowtoenablebloodflow againandthebloodisplacedonthestrip.Theresultisthentaken NormalResults:80-120mg/dlbeforemealsand100/140mg/dlaftermeals 4.Oralglucosetolerancetestisusedtodiagnoseprediabetesanddiabetes.This testisaseriesofbloodglucosemeasurementstakenafteryoudrinkasweet liquidthatcontainsglucose.Thistestiscommonlyusedtodiagnosediabetesthat occursduringpregnancy(gestationaldiabetes).Thistestisnotcommonlyusedto diagnosediabetesinapersonwhoisnotpregnant. Materials/Reagents::FluorideOxalateandotherbloodcollectionequipments, Centrifuge,Insulinkit(NORUDIA® Insulin)Liquidreagent,automatedmachine, glucosesolution Procedure:Patientisaskedtotaketherequiredgram ofsugarsolutionbefore samplecollection.Samplesarecollectedfivetimesat30minintervalsandlastly collectedonce1hrafterthelastsamplecollectionof30minintervals.Theresultis thededucedbythemachines. Results:75gofglucose;fasting92mg/dlormore;1hr180mg/dlormore;2hrs 153mg/dlormore 100gofglucose;morethanorequalto140mg/dl 5.HaemoglobinA1c(alsocalledglycatedhaemoglobin)measureshowmuchsugar (glucose)isstucktoredbloodcells.Thistestcanbeusedtodiagnosediabetes.It alsoshowshowwellyourdiabeteshasbeencontrolledinthelast2to3months andwhetheryourdiabetesmedicineneedstobechanged. Materials/Reagents::FluorideOxalateandotherbloodcollectionequipments, Centrifuge,Glycohemoglobinkit(NORUDIA® HbA1c),automatedmachine, glucosesolution Procedure:SameprocedureasFBS Result:TheresultofyourA1ctestcanbeusedtoestimateyouraverageblood sugarlevel.Thisiscalledyourestimatedaverageglucose,oreAGwhichranges: (126,154,183,212,240,and289)mg/dl 3.4.3 ELECTROLYTESTEST TestOverview:Electrolytesaremineralsfoundinthebodytissuesandbloodin form ofdissolvedsaltswhichhelpstransfernutrientsintobodycellsandwasteof them.Electrolytesalsomaintainahealthywaterbalanceandhelpstabilizebody’s acid/base(pH)level.Themainelectrolytesinthebodyare;Sodium,and

Potassium othersare;CO2-(Bicarbonate),andchloride 3.4.4URICACIDTEST TestOverview:Thisisakidneyorliverfunctiontest,whichmeasurestheamount ofuricacidpresentinabloodsample.Itisproducedfrom thenaturalbreakdown ofbody’scellsandfrom foodeatenandthenfilteredoutbythekidneysand passesoutofthebodyinurinebutiftoomuchisbeingproducedinthebodyor thekidneyisunabletofilterthem normally,itbecomeshighandmaycausesolid crystalsfrom withinjoint,whichmayleadtoapainfulconditioncalledgout.These uriccrystalscanbuildupinjointandnearbytissues,therebyforminghardlumpy depositscalledtophi Results:NormalRange;inmen3.4–7.0mg/dl,inwomen2.4–6.0mg/dl,in children2.0–5.5mg/dl 3.4.5 BLOODUREANITROGENANDCREATININETEST TestOverview:Thistestisusedtodepictthefunctionofkidney.BloodUrea NitrogenandCreatininetestcanbeusedtogethertofindtheBUN-to-Creatinine ratioandwhenthesesubstancesarehighintheblooditmayleadtoheartfailure, dehydration.Bloodureanitrogen(BUN)measurestheamountofNitrogeninyour bloodthatcomesfrom thewasteproducturea.Ureaismadewhenproteinis brokendowninyourbody.Ureaismadeintheliverandpassedoutofyourbodyin urine. NormalResults:Blood Urea Nitrogen (BUN);Adults:10-20mg/dl,Children:5- 8mg/dl BloodCreatinine;Men:0.6-1.2mg/dl,Women:0.5-1.1mg/dl,Children;0.4to 1.0mg/dl 3.4.6 LIVERFUNCTIONTEST Thisisagroupofbloodteststhatdetectsinflammationanddamagetotheliver andalsocheckhowwelltheliverworks.LiverfunctiontestincludesALT(Alanine aminotrans ferase),AST(ASpartateaminotransferaseothersarePT,INR,albumin,bilirubin ALP(Akalinephosphatase) 3.5 MICROBIOLOGYSECTION Microbiologyinvolvesthestudyofmicrobes.Although,microorganismsare generallybeneficialandessentialtolifesomeare,however,pathogenicand causeinfectiousdiseases.Thediagnosticmicrobiologylaboratoryisengagedin theidentificationofinfectiousagents.Theseinfectiousagentsarebroadly classifiedasviruses,bacteria,mycosticagentsandparasites(Protozoansand Helminthes)alsothissectionanalysesbodyfluidsandtissuesforthepresence ofpathogenicmicroorganismsprimarilybymeansofcultureandsensitivity(C&S). ResultsoftheC&Stellthephysicianthetypeoforganismspresentaswellasthe particularantibioticthatwouldbemosteffectivefortreatment 3.5.1 BASICRULESFORWORKINGINTHEMICROBIOLOGYLABORATORY,  Whileworkinginthelaboratory,itisimportantthatyoumustadheretothe followingbasicrules;  Bemethodicalandorderlyinhabits;keeptheworkareacleanespeciallybefore

leavingthelaboratoryanddisinfectantitthoroughlyattheendofday.  Washhandsfrequentlywithsoapandwater  Beforeleavingthelaboratory,placethediscardedglasswareintothe designatedplace.  Culturesarekeptunderincubationandshouldbeinspectedinthemorningand findingsmustbecarefully.  Sendthelaboratoryreportspromptly.Incaseofemergencyaspecialreportis dispatchedorcommunicatedbytelephone. 3.5.2 GENERALREMARKS  Allspecimensforculturemustbecollectedpriortothetherapy.Ifthepatientis onantibiotic,inform themicrobiologistsothathe/shecantakemeasures. Collectthespecimeninadequateamountfrom theinfectioussite.Thisusually instructedbythephysician.  Alwaysusethesterilebottletotransportingthespecimen.  Allspecimenmustbeaccompaniedbyarequestslipwithcompleteinformation, honthepatientname,age,sex,hospitalnumber,sourceofspecimenand clinicalinformationisveryimportantinordertochoosetheappropriatemedium fortheculture.  Enterthedetailsinthelaboratoryregisterandperformeddirectexaminationof thespecimenbeforechoosingthemediafortheculture.  Allcontainersusedforholdingmicrobiologicalspecimensmustbesterilized beforeused.Suchastesttube,culturetubewithandwithoutcap,andplates, containertocollectsputum specimen,bloodspecimenformicrobialculture, penicillinbottlesforcollectionofspinalfluidandotherspecimencontainer (universalbottle)forcollectionofurinespecimenandstoolspecimen. 3.5.3 MATERIALS/APPARATUSUSEDINMICROBIOLOGYSECTION Inoculatingloop,Bunsenburner,Incubator,Weighingbalance,Spatula,Microscopy slide,Coverslip,Stainingrack,Medium plates,Sensitivitydisc,Forcep,Cotton woolswab.

3.5.4 INTRODUCTIONTOLABORATORYGROWTHMEDIA Theseareclassifiedintosixtypes:(1)Basalmedia,(2)Enrichedmedia,(3) Selectivemedia,(4)Indicatormedia,(5)Transportmedia,and(6)Storagemedia. 1.BasalMedia.Basalmediaarethosethatmaybeusedforgrowth(culture)of bacteriathatdonotneedenrichmentofthemedia.Examples:Nutrientbroth, nutrientagarandpeptonewater.StaphylococcusandEnterobacteriaceaegrowin thesemedia. 2.EnrichedMedia.Themediaareenrichedusuallybyaddingblood,serum oregg. Examples:EnrichedmediaarebloodagarandLowenstein-Jensenmedia. Streptococcigrowinbloodagarmedia. 3.SelectiveMedia.Thesemediafavourthegrowthofaparticularbacterium by inhibitingthegrowthofundesiredbacteriaandallowinggrowthofdesirable bacteria.Examples:MacConkeyagar,Lowenstein-Jensenmedia,telluritemedia (Telluriteinhibitsthegrowthofmostofthethroatorganismsexceptdiphtheria bacilli).Antibioticmaybeaddedtoamedium forinhibition. 4.Indicator(Differential)Media.Anindicatorisincludedinthemedium.A particularorganism causeschangeintheindicator,e.g.blood,neutralred,tellurite. Examples:BloodagarandMacConkeyagarareindicatormedia. 5.TransportMedia.Thesemediaareusedwhenspecie-mencannotbecultured soonaftercollection.Examples:Cary-Blairmedium,Amiesmedium,Stuart medium. 6.StorageMedia.Mediausedforstoringthebacteriaforalongperiodoftime. Examples:Eggsalinemedium,chalkcookedmeatbroth.Thesurvivalandgrowth ofmicroorganismsdependonavailableandafavorableconditionenvironment. Culturemediaarenutrientsolutionsusedinlaboratoriestogrowmicroorganisms. Themethodforthepreparationofbasicmicrobiologymediaisgivenbelow. Sterilizationisat121cfor15minutes.pHvaluesare7.0unlessstatedotherwise. 3.5.5 COMMONMEDIAINROUTINEUSE 1.CLEDAgar(CysteineLactoseElectrolyteDeficient):isanon-selectivedifferential platingmedium forthegrowthandenumerationofurinarytractmicroorganism. Preparation:36.0gofmedium issuspendedinoneliterofdistilledwater,slowly heatedandfrequentlystirred.Boiledforaminuteandsterilizedat121OCfor 15minutesandpouredintoPetridishes.Plateswereinvertedwhensolidifiedfor storagepurposesandtoavoidmoisture 2.MacConkeyAgar.Mostcommonlyusedforenterobacteriaceae.Itcontainsagar,

peptone,sodium chloride,bilesalt,lactoseandneutralred.Itisaselectiveand indicatormedium: a)Selective:asbilesaltdoesnotinhibitthegrowthofenterobactericeaebut inhibitsgrowthofmanyotherbacteria. b)Indicator:medium asthecoloniesofbacteriathatfermentlactosetakeapink colourduetoproductionofacid.Acidturnstheindicatorneutralredtopink. Thesebacteriaarecalled'lactosefermenter',e.g.Escherichiacoll.Colourless colonyindicatesthatlactoseisnotfermented,i.e.thebacterium isnon- lactosefermenter,e.g.Salmonella.Shigella,Vibrio. Preparation:50goftheAgarwassuspendedandmeasuredintooneliterof purifiedwaterandmixedthoroughlyandwasheatedwithfrequentagitation, thenboiledforoneminutetocompletelydissolvethepowder.TheAgarwas autoclavedat121OCfor15minutes. 3.ChocolateAgarorHeatedBloodagar.Preparedbyheatingbloodagar.Itisused forcultureofpneumococcus,gonococcus,meningococcusandHaemophilus. Heatingthebloodinactivatesinhibitorofgrowths. Preparation:2litresofdistilledwaterwasaddedto144gofagarpowder. Autoclavingwasdoneat121OCfor15minutesandcooledtill45OCthen5%of defribrinatedbloodwasadded.Heatedslowlyandevenlyto65OC,cooledtill45OC andpouredintoplates 4.BloodAgar.Mostcommonlyusedmedium.5-10%defibrinatedsheepbloodis addedtomeltedagarat45-50°C.Bloodactsasanenrichmentmaterialandalso asanindicator.Certainbacteriawhengrowninbloodagarproducehaemolysis aroundtheircolonies.Certainbacteriaproducenohaemolysis.Typesofchanges: (a)Beta(β)haemolysis.Thecolonyissurroundedbyaclearzoneofcomplete haemolysis,e.g.StreptococcuspyogenesisabetahaemolyticStreptococci. (b)Alpha(α)haemolysis.Thecolonyissurroundedbyazoneofgreenish discolourationduetoformationofbiliverdin,e.g.Viridansstreptococci,and (c)Gamma(γ)haemolysis,or,Nohaemolysis.Thereisnochangeinthemedium surroundingthecolony, 5.NutrientBroth.500gmeat,e.g.oxheartismincedandmixedwith1litrewater. 10gpeptoneand5gsodium chlorideareadded,pHisadjustedto7.3.Uses:(1) Asabasalmediaforthepreparationofothermedia,(2)Tostudysolubleproducts ofbacteria.

3.5.5 DISPOSAL OncethePetridisheshavebeentapedshut,theyshouldnotbeopenedagain.All microorganismsgrownduringtheexperimentshouldbekilledbeforediscarding. Thebestwaytodisposeofbacterialculturesistopressuresterilizethem inaheat stablebiohazardbag.Ifautoclavesorpressurecookersarenotavailableorlarge enoughtomakethisconvenient,analternativeistobleachtheplates.Saturate theplateswitha20%or\"1in5\"householdbleachsolution(inotherwords,1part bleachand4partswater).Letthem sitandsoakovernightinthebleachsolution beforedisposingofthem.Pleasenotethatthebleachsolutioniscorrosiveand needstobethoroughlyremovedafterwards.Inaddition,theplatescanbe incineratedifaccesstoanincineratorisavailable. 3.5.6 PRECAUTIONSTAKENWHENPREPARINGMEDIUM Donottalkwhenpouringmedium onplatesandwhenculturingthesampleon platestoavoidcontaminantsasaresultofunwantedbacterialorenzymes throughsaliva. Thedegreeatwhichweincubateanyculturedsampleisalwaysat37ctoavoid thedeathofthemicroorganism. 3.5.7 GRAM STAINING  Introduction:Inthissection,thestainingofbacteriaasameansfor identificationisdone.BacteriaarebeingidentifiedasGram-NegativeorPositive onthebasisoftheircellwallthicknessafterstaining.Gram positivebacteria holdthedyeandappearpurplewhileGram-Negativebacteriareleasethedye andappearred.  Aim: Toidentifythegram-negativeandthegram positivebacteria.  Apparatus:Stopwatch,Normalsaline,Cleangreasefreemicroscopicslide, Gentianviolet,Lugol’siodine,95%ethylalcohol,Neutralred,Microscope, Sterilizedinoculatingloop. Procedure:Theorganism wasisolatedandsmearedusingthesterilized inoculatingloopinadropofnormalsalineonacleangreasefreemicroscopic slide.ItwaslefttoAir-dry. Thesmearwasplacedonastainingrack,andwasfloodedwithGentianviolet solutionfor30seconds.Itwasrinsedwithwater.Thesmearwasagainflooded withLugol’siodinefor30seconds.Itwasrinsedwithwaterandthesmearwas decolorizedwith95%ethylalcoholfor30seconds,Itwasrinsedwithwater again. Thedecolorizedsmearwascounter-stainedbyfloodingwithneutralredfor30 secondandwasrinsedwithwater.Thebackoftheslidewascleanedwith cottonwoolandallowedtoAir-dry.Theslidewasmountedonthemicroscope afterair-dryingandwasexaminedunder×100immersionobjectives.Theresult wasrecorded. SensitivityTest(Result):Ifthebacteriaaregram positive,apositivesensitive

discisusedwhileanegativesensitivediscisusedforgram negativebacteria. Apurecolonywassub-culturedonaNutrientmedium andsensitivediscwas pickedwiththeaidofasterileforceps,andplacedonthemedium,thenthe platewasincubatedfor24hoursat37OC.Theplatewasreadafter24hoursof incubationtoobservezoneofinhibitionandresistance.Sensitivitytestisalso doneforpathogensthatgrowonthemediabytakingacolonyoftheorganism, streakonthesensitivityagarandaddantibioticsdiscsandincubatefor 24hoursat37OC.

3.5.8 PROCEDUREFORAUTOCLAVING Theinnerelementwasfilledwithwaterandallowedtocoverthesurfaceofthe element,Themedium werearrangedinthesamplebucketandthemachinewas coveredandscrewedtightlybyholdingthescrewoppositeeachotherandthe wingnutwasscrewedtightly. Theswitchonbuttonwasonandalsotheoutletvalvewasopeneddownsoasto increasethetemperatureofthesteam ontheworkload. Itwassterilizedat121⁰Cfor15minutes.Thesafetyvalvewasclosedat120⁰Cand at121⁰C,buttonwasswitchedoff.Themedium wereallowedtocoolfor47⁰C beforepouringinthepetridish PrinciplebehindAutoclaving. Theprinciplebehindthesesterilizationmethodsisbasedonthetemperatureand thetypeofautoclavingoperationperformed.Autoclavingoperationat121⁰Cis referredtoasculturemediaautoclave. 3.6 MICROSCOPY,CULTUREANDSENSITIVITYTESTS Microscopyinvolvestheexaminationofspecimenunderthemicroscope,Culture referstothemicroorganismsthatgrowontheculturemedium afterinoculation andincubationwhilesensitivitytestsdeterminestheantibioticswhichwillbe administeredtothepatients. Theculturedplatesareincubatedfor24hoursat37oCtofacilitatethegrowthof theorganism andchocolateplateswereincubatedinacandlejartofacilitatethe growthofbothaerobicandanaerobicmicrobeswhileotherplateswereincubated aerobically. 3.6.1 STOOLANALYSIS  Introduction:Stoolanalysisinvolvesthecollectionandanalysisoffaecalmatter todiagnosethepresenceorabsenceofamedicalcondition.Duringthe outbreakofcholeraordiarrhea,foodmicrobiologistcollectstoolsampleinto sterileuniversalbottlefrom victimsforfaecalexaminationinthelaboratory.  Aim:Todeterminecystsandovainthestoolofapatient.  Materials/Apparatus:Cleanmicroscopicslide,dropofsaline,

binocularmicroscope,swabstick,coverslip,andaloop.  Procedures:Samplewascollectedintoabottle,adropofnormalsalinewas mountedonacleanmicroscopicslideandmixeditwithasmallportionoffresh faeceswithaloop.Theslidewascoveredwithacoverslipandtheviewed underthelow-powerobjectivebyusing*10and*40forclearviewing.  Results:Theresultswereviewedintwowayswhicharemicroscopicand macroscopic; 1.Macroscopic-----------------Hard,dark,brownstoolformedwithnomucusor bloodseen. 2.Microscopic-----------------PresentofAscarislumbricoides. 3.6.2 URINEMICROSCOPYCULTUREANDSENSITIVITY(M/C/S) Urineformicroscopycultureandsensitivityisanarrayoftestsperformedonurine samplestoexaminethepresenceorabsenceofcellssuchas;epithelialcells,pus cells,redbloodcells,yeastcells,crystals,andbacteria.Itisoneofthemost commonmethodsofmedicaldiagnosis.  Aim:ToidentifyparasitesandBacteriacellintheurineofindividual.  Materials:Microscope,microscopyslide,centrifuge,inoculatingloop,urine sample,medium plates;MacconkeyagarandCLEDagar,Nutrientagar,Gram stainingreagents,Sensitivitydisc,Incubator,sourceofheat.  Procedures: Microscopy:5mloftheurinewastransferredforspinninginabenchcentrifugeat 30000rev/minfor5minutes.Thesedimentwasconcentratedinthetesttubeby decantingoffthesupernatant. Asmalldropofthesedimentwasplacedonacleanslidecoveredwithacoverslip aswetpreparationandthenmountedonamicroscope.Theslidewasexamined usingx40objectivelens. Culture:AloopfulofthesamplewasusedtomakeastreakontheAgarplates. Theplateswerethenincubatedfor24hoursat37OC.Theculturemediawere removedfrom theincubatorafter24hoursandvisiblebacteriagrowthwasread andrecorded. Gram Staining:Primaryandsecondarygram stainingwasdoneonthesmearofa colonyofthebacteria.ThisistoidentifyifthebacteriaisGram positiveorGram negative.  Results: Macroscopy:Thefollowingparametersareexamined 1.Appearance:Clear,turbid,slightlyturbid, 2.Colour:Yellow,straw,Amberyellow,paleyellow. Microscopy:Presenceorabsenceof;Puscells,Epitheliacells,Redbloodcells, Yeastcells,BacteriacellsandCrystalscanbeseeninthewetpreparation. Culture:ThefollowingBacteriacanbeisolatedfrom Urinesamples;Klebsiellaspp, Staphylococcusaureus,Coniforms

3.6.3 EYE,EARNOSEANDTHROAT,WOUNDANDPUSSWAB  Procedure:Thesesamplesarecollectedusingadrysterilecottonwoolswab; theyaretheninoculatedintoChocolateandMacconkeyagar.Gram stainingis thencarriedoutoneachspecimen.Pathogensthatarelikelytobeobserved include;  WoundandPusswab:Staphylococcusaureus,Sreptococcuspyogenes, Clostridium perfrigens  Eyeswab:Haemophilusinfluenza,Pseudomonasaeruginosa,Betahaemolytic streptococcusetc.  Earswab:Escherichiacoli,Proteusspetc.  ThroatSwab:ThroatculturesaresubmittedprimarilyforthedetectionofGroupA Streptococcus.Whenobtainingthespecimen,depressthetonguewithatongue blade,andswabthetonsillarpillarsandbehindtheuvulaincludinganyinflamedor purulentsitesAvoidtouchingthetongue,cheeksorteeth.Immediatelyplacethe swabbackintotheculturettesleeveandcrushtheampule 3.6.4 HIGHVAGINAL,URETHRALANDENDOCERVICALSWAB.  TestOverview:Thisisusetodetectthecausativeorganismsoffemale reproductivesystem infectionsandtheirsensitivitytoantibiotics. Procedure:Aswabstickwasusedtocollectspecimenfrom theaffectedarea andtheninoculatedintosterilemediawhichincludeChocolateandMacConkey agar.Theseplateswereincubatedat37cfor24hoursandwereexaminedfor anypathogenicgrowth,ifthereisanygrowththenasensitivitydiscisplacedon apurecultureofisolate. Gram stainingisthencarriedoutoneachspecimen,awetpreparationofthe swabcanbemadebydroppingnormalsalineintotheswabcontainerandthe swabstickisrubbedonaslidecoveredwithaslipandviewedunderthe microscope;puscells,epithelialcells,yeastcellsetccanbeviewed.  Result:Possiblepathogensinclude;Neisseriagonorrhea,Trichomonasvaginals, Candidaspp,Clostridium perfrigensetc. 3.6.5 MANTOUXSKINTEST  Introduction:TheMantouxtestorMendel-Mantouxtest(alsoknownasthe tuberculinsensitivitytest,orPPDtestforpurifiedproteinderivative)isa screeningtoolfortuberculosis(TB)Itisoneofthemajortuberculinskintests usedaroundtheworld,largelyreplacingmultiple-puncturetestssuchasthe Tinetest.Tuberculinisaglycerolextractofthetuberclebacillus.Purified proteinderivative(PPD)tuberculinisaprecipitateofspecies-nonspecific moleculesobtainedfrom filtratesofsterilized,concentratedcultures.  Material/Equipments:Millimeterrule,tuberculininjection,Personalprotective equipment,cottonwool.  Procedures:Astandarddoseis5tuberculinunits(TU-0.1ml)isinjected intradermally(betweenthelayersofdermis)andread48to72hourslater.This

intradermalinjectionistermedtheMantouxtechnique.Apersonwhohasbeen exposedtothebacteriaisexpectedtomountanimmuneresponseintheskin containingthebacterialproteins.Ifapersonhashadahistoryofapositive tuberculinskintest,orhadarecenttuberculinskintest(withinoneyear), anotherskintestshouldbeused  Result:Withintwodaysofinjection,thereactionisreadbymeasuringthe diameterofinduration(palpableraised,hardenedarea)acrosstheforearm (perpendiculartothelongaxis)inmillimeters.Ifthereisnoinduration,the resultshouldberecordedas0mm.Erythema(redness)shouldnotbemeasured. Atuberculintestconversionisdefinedasanincreaseof10mm ormorewithin atwo-dayperiod,regardlessofage.Alternativecriteriaincludeincreasesof6, 12,15or18mm.

CHAPTERFOUR 4.0 SUMMARY,CHALLENGESENCOUNTERED,ANDCONCLUSION RECOMMENDATION 4.1 SUMMARYOFATTACHMENTACTIVITIES DuringmyperiodattheLagosStateUniversityHealthCenterasaSIWES student,catalogingsomeinformationmaterialsforthelaboratoryandIalsodid someactivitiesatthereceptionsuchas:attendingtopatients,confirmingand examiningtheirrequestforms,enteringtheirdetailsintotheregister,detailing them concerningthetesttheyaretoundergoanddirectingthem towhereisto becarriedout. Iwaslatertransferredtothelaboratoryandwasintroducedtovarious departmentsandthesafetyprecautionsandtestscarriedoutineach department. 4.2 CHALLENGESENCOUNTERED Themainproblemsencounteredweregettingplacementandtransportation. Itwasquitechallengingformethatliveinfarplacetogettotheorganisation everyworkingday.Iwasnotgivenanyremunerationorallowance,other problemsencounteredduringthetrainingwasattendingtodifferentpeoplewith differentpersonalitiesatthereception. 4.3 CONCLUSION MythreemonthsindustrialattachmentwithLagosStateUniversityHealth Centerhasbeenoneofthemostinteresting,productive,instructiveand educativeexperienceinmylife.Throughthistraining,Ihavegainednewinsight andmorecomprehensiveunderstandingabouttherealindustrialworking conditionandpracticeandalsoimprovedmysoftandfunctionalskills. AllthesevaluableexperiencesandknowledgethatIhavegainedwerenot onlyacquiredthroughthedirectinvolvementintaskbutalsothroughother aspectsofthetrainingsuchas:workobservation,supervision,interactionwith colleagues,supervisors,superiorandotherpeoplerelatedtothefield.Italso exposedmetosomecertainthingsaboutmedicalenvironment.Andfrom what Ihaveundergone,Iam surethattheindustrialtrainingprogrammehasachieved itsprimaryobjective. Asaresultoftheprogramme,Iam nowmoreconfidenttobuildmyfuture careerwhichIhavealreadystartedwithLagosStateUniversityHealthCenter 4.4 RECOMMENDATION IrecommendthatallinstitutionsorbodiesinvolveinStudentIndustrial WorkingExperienceScheme,shouldprovideplacesforindustrialattachment forStudentIndustrialTrainingFundandalsopaysomeallowancestostudents andthecompanyshouldprovidemoresafetyequipmentstopreventfurther environmentalandhealthhazards. Also,tostudentsthataretoundergothetraining,Irecommendthatthey shouldtakeitveryseriously,becauseitisoneofthemostimportantpartsof theirstudieswhichwillhelpthem buildaverysignificantandeffectivemeaning

intheircareerpursuit.