試驗(yàn)室檢測及數(shù)據(jù)分析應(yīng)用課件

InterpretationofSerologicalresults

血清結(jié)果的分析Dra.BeatrizCardoso,MSc,ACPVDr.OscarMorales,ACPVLohmannAnimalHealthSerology血清學(xué)Whatisserology?什么是血清學(xué)？Whatisitfor?

血清學(xué)的意義Howtogetthebestserologicalresults?

怎樣獲得最佳的血清結(jié)果？Howtointerpretserology?broilersLayersandbreeders怎樣分析血清學(xué)結(jié)果？－肉雞－蛋雞和種雞Samplesandserumbanks

采樣和血清保存Whatisserology?

什么是血清學(xué)？Detectionofantibodies

抗體的檢測QualitativeTests定性試驗(yàn)Answerisyesorno

回答“是”或者“不是”Resultsare“positive”or“negativo”

結(jié)果為“陽性”或“陰性”RapidPlateAgglutinationtest

快速平板凝集試驗(yàn)AgarGelPrecipitationtest

瓊脂凝膠沉淀試驗(yàn)Semi–Quantitativetests

半定量試驗(yàn)Resultsexpressedintitres

結(jié)果用滴度表達(dá)Basedonserialdilution

基于連續(xù)稀釋Resultsexpressed

in2x,10x(2log,10log)

結(jié)果表達(dá)為2x,10x(2log,10log)HI,Serumneutralization

HI,血清中和試驗(yàn)Quantitativetests定量試驗(yàn)Resultsinabsolutenumbers

結(jié)果用絕對數(shù)表示Resultisthecolorimetricreactionofthesubstrateproportionaltothequantitiyofantibodies

結(jié)果是與抗體數(shù)量成比例的物質(zhì)的比色反應(yīng)ELISA

酶聯(lián)免疫吸附試驗(yàn)Sensibility敏感性Abilitytodetermineminimalquantitiesofantibodies

敏感性是指能檢測到的最小抗體的數(shù)量Avoidfalsenegatives

避免假陰性Falsepositives假陽性Diseasecontrolanderradication

用于疾病控制和凈化Example:SPA

舉例：SPAIdeal理想狀況Calculations計(jì)算Especificity:特異性RealNegatives/TotalnumberInfectedx100真正的陰性/合計(jì)感染數(shù)x100specificity=[80/(80+20)]x100=80%Aplicationinthefield現(xiàn)場運(yùn)用Farmwith10.000birds某場10000羽雞100%checked100％進(jìn)行檢測500positives出現(xiàn)500陽性490truepositives10falsepositives490真陽性10假陽性specifictest特異性試驗(yàn)Calculations2計(jì)算2Sensibility=80/(100)x100=80%

敏感性＝80/(100)x100=80%specificity=100/(100)x100=100%

特異性＝100/(100)x100=100%specificityandSensibility

特異性和敏感性Typeofmethod

方法的類型Antigenused使用的抗原Conditionsofthetechnique

技術(shù)條件Testcommonlyusedinpoultry

家禽常用的檢測方法RPA-- Mycoplasma,Salmonella

AGP-- IBV,IBD,Marek,Reo

HI-- ND,IBV,EDS,Mycoplasma,Coryza

Serology血清學(xué)Diagnostics

診斷Monitoring

監(jiān)測INTERPRETATION

分析Criteriaforinterpretation

分析的標(biāo)準(zhǔn)WhatdiseasedoIwanttoverify?

我要驗(yàn)證什么?。縁orwhatpurpose?DiagnosticsorMonitoring?

分析的目的是什么？診斷或監(jiān)測？WhattypeofexploitationdoIhave?

我該采取什么方法？HowshouldIinterprettheresults

我該怎樣分析結(jié)果？Basisfortheinterpretation

分析的基礎(chǔ)PresenceofantibodiesInfectionDevelopmentoftheimmunity

存在抗體－感染－免疫力的產(chǎn)生AbsenceofantibodiesThereisnoinfectionTheInfectionwasveryrecentTheimmunityisinanearlystageofdevelopment沒有抗體：－沒有感染－最近剛感染－免疫力正在產(chǎn)生早期階段Interpretation:qualitativetests

分析：定性試驗(yàn)Negative陰性PositivoSample樣品No

infection

未感染Acutephase

急性期Lowlevelof

antibodies

抗體水平太低Re-test2weekslater

2周后重新檢測Useamoresensitivetest使用更敏感的試驗(yàn)Vaccine疫苗live–no

challenge活苗－未攻毒killed–vaccinationfailure滅活苗－免疫失敗Challengetest--verifyprotection

攻毒試驗(yàn)－驗(yàn)證保護(hù)力Takesamplesatalatermoment過一段時(shí)間再次采樣Interpretation:qualitativetests

分析：定性試驗(yàn)NegativoPositive陽性Sample陽性Infection感染Sero-responsetotheantigen

對抗原血清學(xué)反應(yīng)Confirmation確診Useatestwithahigherspecificity

使用敏感性更高的試驗(yàn)Isolationoftheagent病原分離Vaccine疫苗live–fieldchallengeorvaccineresponse活毒－野毒感染或疫苗反應(yīng)killed–vaccineresponse滅活苗－疫苗反應(yīng)Interpretation:quantitativetests

分析：定量試驗(yàn)NegativoHightitre高滴度Sample樣品Vaccine疫苗live–fieldchallengeorvaccineresponse活毒－野毒感染或疫苗反應(yīng)killed–vaccineresponse滅活苗－疫苗反應(yīng)Confirmation確診Useatestwithahigherspecificity

使用敏感性更高的試驗(yàn)Isolationoftheagent病原分離Infection感染Sero-responsetotheantigen

對抗原血清學(xué)反應(yīng)Vaccine疫苗GoodresponsetothevaccinationGoodvaccinecoverage接種后的良好反應(yīng)Interpretation:(semi-)quantitativetests

分析：半定量試驗(yàn)LowCV低CVNarrowdistributionoftitres

狹窄的滴度分布No

infection沒有感染ChronicInfectioninthepopulation雞群內(nèi)慢性感染SamplesInterpretationinbroilers

對肉雞群的分析LevelsofMaternallyDerivedAntibodies

母源抗體的水平KnowledgeaboutthesanitarystatusOutbreaks.Infectionsorchallenge

關(guān)于衛(wèi)生狀況的知識－疾病爆發(fā)、感染、攻毒CheckthevaccinationprogrammesFixeddates檢查免疫程序固定日期NormalCurve

正態(tài)曲線Comparingcurves

曲線比較Interpretation–chickensorlivevaccines

分析－雛雞或活苗血清學(xué)結(jié)果外表性能分析措施Lowtitre+CVgood+goodperformance

低滴度＋良好的CV＋良好的性能=continuewiththecontrolprogramme繼續(xù)控制程序HighTitre+goodperformance

高滴度＋良好的性能=continuethecontrolprogrammeandKeepaneyeonthebiosecurity!繼續(xù)控制程序，注意生物安全！Watchout!注意Vaccinatedflock免疫過的雞群=Infectionwithprotection

具保護(hù)力的感染=Vaccinationwithahotstrain

用強(qiáng)毒接種Non-vaccinatedflock未免疫的雞群=pastinfection

過去感染過=Infectionwithamildstrain

弱毒株感染過Hightitre+narrowCV

高滴度＋窄CVOldinfectionGoodvaccinecoveragewiththeuseofastrongvaccinestrain使用了強(qiáng)的疫苗毒株，具有良好的疫苗覆蓋性Hightitre+wideCV

高滴度＋寬CVAcuteinfection急性感染Poorvaccinecoverageusingahotvaccinestrain

使用了強(qiáng)毒疫苗，疫苗覆蓋性差。Hightitre+poorperformance

高滴度＋差性能Isolateandidentifytheagent.分離鑒定病毒Improvethecontrolprogramme.改進(jìn)控制程序Improvethebiosecurity改進(jìn)生物安全。=fieldinfection

Challengeanddisease野毒感染和疾病Lowtitre+poorperformance

低滴度＋差性能acuteinfection–noantibodiesyet

急性感染－還未產(chǎn)生抗體otherproblems其它問題Retest1weeklater

1周后重新檢測Lookforotherpossiblecause

尋找其它可能的問題InterpretationbyAgent

按病原分析Interpretationinbreedersorlayers

種雞和蛋雞的分析KnowledgeofthesanitarystatusOutbreaks,infectionorchallenge?衛(wèi)生狀況的知識－爆發(fā)、感染、攻毒CheckvaccinationprogrammesPre-fixeddates－檢查免疫程序Comparisonwithproductioncurves

比較生產(chǎn)曲線Highandlowtitres-howmuch?ItisallisamatterofperspectiveHighorlowtitres-howhigh?Howlow?抗體滴度高和低的認(rèn)識Baselines基準(zhǔn)線Highorlowtitres-cuanto?滴度高或低－多少？Basededatos數(shù)據(jù)的基準(zhǔn)線Curves-IBD(Real)

IBD的抗體滴度曲線Serologyoftheindividual

個(gè)體血清學(xué)Serologyofpopulations(flocks)群體血清學(xué)Samples采樣數(shù)量Baseline-minimum23,ideal28

基準(zhǔn)線－最低23份，理想28份

Monitoring-minimum15

監(jiān)測－最低15份Diagnostics-between28and146

診斷－（28～146）份Serum“banks”血清保存Research研究PruebascomprobatoriasImportantfordiagnostics診斷的重要性Security安全性Adequatestorageoftheserum–avoidevaporation

正確存儲血清，避免蒸發(fā)Collectingsamples采集樣品Monitoring監(jiān)測Consistence

時(shí)間安排VaccinationProgrammes

免疫程序Mineral-oilvaccines

礦物油疫苗Feed飼料Diagnostics診斷Pairedsamples雙份采樣SerotecaConditionsunderwhichthesamplesaretaken