SpecializedLaboratoryServices

AnatomicPathology

TheAnatomicPathologyLaboratoryconsistsofthefollowing: Cytopathology,ElectronMicroscopy,Histology, Immunofluorescence/Immunohistochemistry, andFluorescent InSituHybridization(FISH)Laboratories.

•Cytopathology

TheCytologyLaboratoryperformscytologic examinationofgynecologicandnon-gynecologic specimens.Ourfine-needleaspirationserviceincludes anoutpatientclinicforreferredpatientswithsuperficial

andpalpablelesions.Thelaboratoryutilizesstate-of-the- artliquid-basedmonolayertechnologypreparationfor bothgynecologicandnon-gynecologicspecimens.

•ElectronMicroscopy

TheElectronMicroscopyLaboratoryprovidesa further diagnostictoolwithidentificationofultrastructural features.

•Histology

TheHistologyLaboratoryperformsa widevarietyof routinehistotechnologicprocedures,histochemical stains,andinsituhybridizationprobes.

•Immunofluorescence/Immunohistochemistry

TheImmunohistochemistryLaboratoryperformsover

100antibodiesonbothtissuesectionsandcytology specimens.

•FluorescentInSituHybridization(FISH)Laboratory TheFISHlaboratoryperformsfluorescentinsitu hybridizationonparaffinembeddedtissueandsome cytologyspecimens.

BloodBank

TheBloodBankLaboratoryprovidesbloodtypeandscreen testingandafullrangeofantibodyidentificationservices includingpanels,adsorptions,directantiglobulin(Coombs) tests,eluates,titers,andphenotyping.TheBloodBankprovides

70,000bloodproductsayearfortransfusion.

Chemistry

TheChemistryLaboratoryat Barnes-JewishHospitalismade upof6areas:SpecimenReceivingProcessing,General Chemistry,SpecialChemistry,ToxicologyTherapeuticDrug Monitoring,QualityAssurance,andAssayDevelopment.

•SpecimenReceivingProcessing

TheReceivingProcessingareareceivesandsorts patientspecimenstobeperformedinChemistry. Specimensareprocessedon2 fullyautomatedsystems capableofidentifying,centrifuging,aliquotingand sortingspecimensforrapiddeliverytothetestingareas.

•GeneralChemistry

TheGeneralChemistryareaperformsover2 million STATandroutineprofilesandindividualtestsannually onstate-of-the-artequipment.Thecriticalcarecardiac workstationperformsatroponinassayontheaverageof

1 every8minutes.Ourmobilebloodgas/chemistries unitisoperatedintheSouthCampustransplantation operatingroomtosupportlivertransplantprocedures.

•SpecialChemistry

TheSpecialChemistryareaperformsmanyspecial hormoneandtumormarkerassaysalongwiththyroid functiontests,someserologicaltests,andmanual chemistries.STATaswellasroutinetestingis provided formanyoftheSpecialChemistryanalytes.Special Intraoperativeparathyrpidhormone(PTH)analysisis offeredtosupportparathyroidectomysurgeriesonboth northandsouthcampuses.

•ToxicologyTherapeuticDrugMonitoring

TestingforbothDrugsofAbuseandtherapeuticdrugsis performedinToxicology.Instrumentationincludes automatedimmunoassayanalyzers,liquid chromatography-tandemmassspectrometry(LC-MS/ MS),gaschromatography(GS),andthinlayer chromatography(TLC).BothSTATandroutinetesting is offered.

•QualityAssurance

TheQualityAssuranceareaprovidesmonitoringof qualitycontrolandtheQualityImprovementprogram utilizingasophisticatedcomputerizeddatasystem.They alsoprovidein-housecontinuingeducationforthe laboratorystaffandsupporttheSaintLouisUniversity DoisyCollegeofHealthSciences,Departmentof ClinicalLaboratoryScience.

•AssayDevelopment

TheAssayDevelopmentareainvestigatesnew methodologiesandinstrumentationforthelaboratory. WorkingcloselywithWashingtonUniversityMedical Directorsandthefield’sleadingmanufacturers,the developmenttechnologistsmaintaintheBarnes-Jewish Hospitalreputationasaleaderinclinicalchemistry research.

Hematology

TheHematologyLaboratoryisresponsibleformeasurementof hematologicandhemostaticparametersaswellasurinalysis andflowcytometry.

•Hematology

Employsafullyrobotichematologysystemcapableof processingthousandsofspecimensa dayproviding completebloodcellcounts,reticulocytecounts,and5- partdifferentialsaswellaspreparingandstainingslides.

•Manualbenchtests

Includecellcountsonbodyfluids,erythrocyte sedimentationratesandcrystalanalysisonjointfluids.

•Hemoglobinopathyevaluation

EvaluationbyHPLCandelectrophoreticanalysisof hemoglobinchains

•Hemostasis/thrombosis

—Comprehensiveanalysisofcoagulationfactorsand inhibitors

—EvaluationforvonWillebranddisease

—Evaluationofhypercoagulabilitysyndromes

—Lupusanticoagulanttesting

•Plateletfunctiontesting

—PlateletfunctionscreeningwithPFA-100 instrument

—Plateletaggregationwithmultipleagonists

performedonplateletrichplasma

•Urinalysis

—Automatedurinechemistry

—Automatedandmanualurinesedimentmicroscopy

•FlowCytometry

TheFlowCytometryLaboratoryperformsassaysfor clinicaldiagnostictesting:T-cellandB-cellquantitation forimmunestatus,leukemia/lymphomaimmunopheno- typing,fetalhemoglobins,CD34enumeration,and paroxysmalnocturnalhemoglobinuria(PNH).

HLATissueTyping

TheHLATissueTypingLaboratoryprovides:

•Histocompatibilitytyping,matchingandantibody screeningforrenal,hepatic,cardiac,pancreasand pulmonarytransplantation.

•DNAtypingforrelatedandunrelatedbonemarrow transplantation.

•Singledonorapheresiscomponenttherapyforpatients withhematologicdisorders.

•ClassI andClassII phenotypingfordiseaseassociation studiesincludingHLA-B27testing.

ClassItypingconsistsoftypingforHLA-A,-B,-Callelesand

ClassIIforHLA-DRandDQalleles.

DNAtypingis performedbythereversedot-blot,sequence- specificprimertechniques,andsequence-basedtypingfor HLA-A,B,C;DRB1;DRB3;DRB4;DRB5;andDQB1

Immunology

TheClinicalImmunologyLaboratoryperformsdiagnostic testingin4 majorareas:serologicmarkersofautoimmune disease,analysisofthecomplementsystem,serodiagnosisof infectiousdisease,andspecialproteinstudiesformonoclonal proteindetection.TheClinicalImmunologyLaboratory welcomesquestionsandconsultationsconcerningallaspectsof clinicaldiagnosticimmunology.

•Serologicmarkersofautoimmunediseaseinclude:

—Antinuclearantigensbyimmunofluorescence

—Anti-DNAbyimmunofluorescenceand radioimmunoassay

—Extractablenuclearantigensandanticardiolipin antibodiesbyenzyme-linkedimmunosorbentassay (ELISA)

—Rheumatoidfactor

—Antimitochondrial,andsmoothmuscleantibodies byimmunofluorescence

•Analysisofthecomplementsystemincludes:

—Determinationoftotalhemolyticcomplement

—C3,C4,andC1esteraseinhibitorlevelsby nephelometry

•Serodiagnosisofinfectiousdiseaseinclude:

—Immunestatustodetermineexposureby immunofluorescence,agglutination,orELISA

—Syphilistestingbynontreponemal(RPRand

VDRL)andspecifictreponemal(FTA)tests

•Specialproteinstudiesperformedinclude:

—Serumandurineproteinelectrophoresis

—Immunofixationofallabnormalserumandurine specimens

—Cryoglobulinstudiesonserum

MetabolicGenetics

Barnes-JewishHospitalLaboratoryreferstestingforgenetic metabolicdiseasestotheMetabolicGeneticsLaboratoryatSt. LouisChildren’sHospital.Thelaboratoryofferstestsdesigned toscreenforarangeofgeneticmetabolicdiseases.Aminoacid andorganicanalysisarefrequentlyperformedtogetheron specimensobtainedfromchildrensuspectedofhavinganyof dozensofinbornerrorsofaminoacid,organicacid,fattyacid, andcarbohydratemetabolismdisorders.Aminoacidsare analyzedbyhigh-performanceliquidchromatography,yielding quantitativeresultson42constituents.Organicacidsare analyzedbygaschromatography-massspectroscopy(GC-MS) yieldingsemiquantitativeresultsonanyofdozensofpotentially diagnosticallyimportantconstituentswhichmightbefoundin abnormalamounts.

Othertestsperformedbythislaboratoryincludevolatile organicacids(propionicandisovalericacids)analysis.

Thelaboratoryhasbeenactivelyinvolvedinresearchand developmentofnewtechnologiesofspecialinterestinpediatric medicine.Theseincludethedevelopmentofibuprofen measurementbyGC-MSandassessmentofsensitivityofurine aminoacidanalysisfordetectionofhyperoxaluria.

Microbiology

TheMicrobiologyLaboratoryisresponsibleforisolationand identificationofbacteria,mycobacteria,fungi,andparasites fromspecimens,aswellassusceptibilityandantigentesting. MoleculardetectionofChlamydiatrachomatisandNeisseria gonorrhoeae,moleculartypingofbacteria,andmolecular epidemiologyservices(bacterialstraintyping)areperformedin theMicrobiologyLaboratory.Virologyisperformedat St. LouisChildren’sHospitalLaboratory.See“Virology”(this section).

MolecularDiagnostics

A broadselectionoftestsisavailablein3 generalcategories:

•MolecularOncology

•MolecularGenetics

•Pharmacogenetics

Alltestsarebasedondetectionofspecificgeneticvariations usingisolatedDNAorRNA.Avarietyofclinicalspecimens maybeevaluateddependingonthedetectionmethodused, includingfresh,frozenandarchivaltissues.

Examplesofthesetestsincludethefollowing:

•Beckwith-Wiedemannsyndrome-abnormal

methylation

•BCR-ABLt(9;22) p210 by IS

•Cysticfibrosis

•FactorVLeidenmutationanalysis

•FragileXsyndromeandfragilex-associatedtremorand ataxia

•IGHgene rearrangements

•IGHhypermutationstatus

•NPM1Nucleophosmin gene mutation

•Prader-WilliandAngelmansyndromes-abnormal methylation

•Prothrombinmutationanalysis

•PML-RAR-alphat(15;17)inacutepromyelocytic leukemia

•Monitoringofhematopoieticstemcelltransplant

engraftment–STRmarkers

•Hydatidiformmoleverification–STRmarkers

•TCRG gene rearrangements

•UGT1A1mutationdetection-Irinotecantoxicity

Fordetailedinformationonthesetestsandmore,seethe MolecularDiagnosticLaboratorywebsiteavailableat URL: pathology.wustl.edu/patientcare/moldiagnostic.php

PheresisCenter

ThePheresisCenterprovidesservicesasbothaninpatientand outpatientcenter.Specificfeaturesinclude:

•Singledonorplateletcollection-fromadedicated volunteerdonorpopulation,at2sites

•Wholebloodcollections

—Autologousdonations-requiringordersfor collectionfromtheattendingphysician

•Manualtherapeuticphlebotomy-forthetreatmentof specifichematologicdisordersasprescribedbythe attendingphysician

•Therapeuticpheresis- usingstate-of-the-artbloodcell separatorsinthetreatmentofvariousdiseaseprocesses

•Peripheralbloodstemcellcollections-forpatients undergoingbonemarrowtransplantinconjunctionwith theBarnes-JewishHospitalBoneMarrowTransplant Program

•Photopheresis-forpatientsrequiring immunomodulationforcutaneousT-celllymphoma, scleroderma,hearttransplantrejection,lungtransplant rejection,orgraft-versus-hostdisease.

Point-of-CareTesting

Thepoint-of-caretesting(POCT)teammaintainsresponsibility fortheauthorizationandoversightofanylaboratorytesting doneoutsidetheDepartmentofLaboratorieswithinBarnes- JewishHospital,ie,point-of-caretesting,nearpatienttesting, bedsidetesting.TheprogramisgovernedbytheClinical LaboratoryImprovementAmendments,theCollegeof AmericanPathologists,theMissouriDepartmentofHealthand SeniorServices,andtheJointCommissiononAccreditationof HealthcareOrganizations.

Approvedpoint-of-caretestingmustprovidea clearbenefitto patientcarewithrespecttomanagementofdiagnosis, treatment,prevention,and/orsafetyoverconventional laboratorytesting.AuthorizationtoperformPOCTisgrantedto physician,nursing,andotherqualifiedpersonnelwhohave beensuccessfullytrainedandwhoannuallysuccessfully completecompetencytestingforthe specifiedprocedure. POCTwithinBarnes-JewishHospitalincludeswaivedand moderatelycomplextesting.POCTisonlyperformedusing instruments,testkits,andtechnologiesapprovedbythe DepartmentofLaboratoriesandtheWashingtonUniversity DepartmentofPathologyImmunology,Divisionof

LaboratoryMedicine.RefertothePoint-of-CareTesting (POCT)ApprovedTestingCatalog,DT.01.01.01,forcomplete listofapprovedpoint-of-caretests,locations,instruments,test kits,andmethods.

Virology

The Virology Laboratory at St. Louis Children’s Hospital offers a full range of testing for common viral agents isolated from adult and pediatric patients. Cell culture and shell-vial assays are the mainstay of offered tests. Specimens are received, processed, and inoculated into a variety of cell cultures which support the growth of common viral isolates. Inoculated cell cultures are then observed daily for the development of viral cytopathic effect. In an effort to provide a more timely diagnosis, the laboratory also offers rapid antigen detection tests for a variety of viral agents. These rapid assays rely on the use of monoclonal antibodies in fluorescent antibody test format to detect viral antigens in cells present in the specimen. Viruses detected in these rapid assays include: respiratory syncytial virus (RSV), influenza virus, parainfluenza viruses, adenovirus, and varicella-zoster virus (VZV). Recognizing the increased usage of antiviral drugs and concomitant development of resistance to these agents, the Virology Laboratory also offers susceptibility testing for isolates of herpes simplex virus (HSV) and cytomegalovirus (CMV) to the drugs acyclovir and ganciclovir. The laboratory also performs cell culture for Chlamydia.

The Molecular Virology section performs tests using PCR- based technology to specifically amplify and detect the presence of nucleic acid sequences from a variety of infectious disease agents in patient specimens.

Agents detected include the following organisms causing systemic infections: CMV, Epstein-Barr virus (EBV), human herpes virus type 6 (HHV-6), parvovirus B19, and Ehrlichia species. Blood is the specimen of choice for these agents. The laboratory also offers a test for the detection of Bartonellahenselae the causative agent of another systemic illness, cat- scratch disease. For this agent, the specimen of choice is either lymph node tissue or lymph node aspirate. A second group of agents include those associated with infections of the central nervous system. These agents include: HSV, CMV, VZV, EBV, HHV-6, human polyomavirus JC, enteroviruses, and Toxoplasma gondii. The specimen of choice for these infections is spinal fluid. To rule out renal infection with the human polyomavirus BK, urine or blood specimens are tested. Lastly, testing for bacterial agents of whooping cough, Bordetella pertussis/Bordetellaparapertussis is accomplished by testing nasopharyngeal swab specimens submitted in saline.