试验室检测及数据分析应用讲稿.ppt

关于试验室检测及数据分析应用第一页，讲稿共七十三页哦Serology血清学What is serology?什么是血清学？What is it for?血清学的意义How to get the best serological results?怎样获得最佳的血清结果？How to interpret serology?broilersLayers and breeders怎样分析血清学结果？肉鸡蛋鸡和种鸡Samples and serum banks 采样和血清保存第二页，讲稿共七十三页哦What is serology?什么是血清学？Detection of antibodies 抗体的检测第三页，讲稿共七十三页哦Types of antibodies抗体的类型IgAIgMIgEIgG第四页，讲稿共七十三页哦Qualitative Tests定性试验Answer is yes or no 回答“是”或者“不是”Results are“positive”or“negativo”结果为“阳性”或“阴性”Rapid Plate Agglutination test 快速平板凝集试验Agar Gel Precipitation test 琼脂凝胶沉淀试验第五页，讲稿共七十三页哦Semi Quantitative tests半定量试验Results expressed in titres 结果用滴度表达Based on serial dilution 基于连续稀释Results expressed in 2x,10 x(2log,10log)结果表达为2x,10 x(2log,10log)HI,Serum neutralization HI,血清中和试验第六页，讲稿共七十三页哦Quantitative tests定量试验Results in absolute numbers 结果用绝对数表示Result is the colorimetric reaction of the substrate proportional to the quantitiy of antibodies 结果是与抗体数量成比例的物质的比色反应ELISA 酶联免疫吸附试验第七页，讲稿共七十三页哦Sensibility敏感性Ability to determine minimal quantities of antibodies 敏感性是指能检测到的最小抗体的数量Avoid false negatives 避免假阴性False positives 假阳性Disease control and erradication 用于疾病控制和净化Example:SPA 举例：SPA第八页，讲稿共七十三页哦Specificity特异性Antibodies developed against a specific agent 由特异性抗原产生的抗体Avoid false positives 避免假阳性False negatives 假阴性Example:Imnmunofluorescence 举例：免疫荧光试验第九页，讲稿共七十三页哦Ideal理想状况第十页，讲稿共七十三页哦Calculations计算Sensibility:敏感性Real Positives/(Total number Infected)x 100真正的阳性/(合计感染数)x 100Sensibility=98/(98+2)x100=98%第十一页，讲稿共七十三页哦Calculations计算Especificity:特异性Real Negatives/Total number Infected x 100真正的阴性/合计感染数x 100specificity=80/(80+20)x100=80%第十二页，讲稿共七十三页哦Calculations计算 Sensibility=98%specificity=80%敏感性98特异性80第十三页，讲稿共七十三页哦Aplication in the field现场运用Farm with 10.000 birds某场10000羽鸡100%checked100进行检测500 positives出现500阳性490 true positives10 false positives490真阳性 10假阳性第十四页，讲稿共七十三页哦specificity and Sensibility特异性和敏感性Type of method 方法的类型Antigen used 使用的抗原Conditions of the technique 技术条件第十七页，讲稿共七十三页哦Test commonly used in poultry家禽常用的检测方法RPA-Mycoplasma,SalmonellaAGP-IBV,IBD,Marek,ReoHI-ND,IBV,EDS,Mycoplasma,Coryza第十八页，讲稿共七十三页哦Test commonly used in poultry家禽常用的检测方法SN-IBV,IBD,all for research purposesELISA nearly all agents第十九页，讲稿共七十三页哦Serology血清学Diagnostics 诊断Monitoring 监测INTERPRETATION 分析第二十页，讲稿共七十三页哦Dogma o Paradigma?USUALLY THE HIGHER THE TITRE,THE BETTER THE PROTECTION通常滴度越高，保护力越高通常滴度越高，保护力越高第二十二页，讲稿共七十三页哦Basis for the interpretation分析的基础Presence of antibodiesInfectionDevelopment of the immunity 存在抗体 感染 免疫力的产生Absence of antibodiesThere is no infectionThe Infection was very recentThe immunity is in an early stage of development没有抗体：没有感染最近刚感染免疫力正在产生早期阶段第二十三页，讲稿共七十三页哦Interpretation:qualitative tests分析：定性试验Negative阴性PositivoSample样品No infection 未感染Acute phase 急性期Low level of antibodies 抗体水平太低Re-test 2 weeks later 2周后重新检测Use a more sensitive test使用更敏感的试验Vaccine疫苗live no challenge活苗未攻毒killed vaccination failure灭活苗免疫失败Challenge test-verify protection 攻毒试验验证保护力Take samples at a later moment过一段时间再次采样第二十四页，讲稿共七十三页哦Interpretation:(semi-)quantitative tests分析：半定量试验Negative or low titre阴性或低滴度PositivoSample样品No infection 未感染Acute phase 急性期Low level of antibodies 抗体水平太低Re-test 2 weeks later 2周后重新检测Use a more sensitive test使用更敏感的试验Vaccine疫苗live no challenge活苗未攻毒killed vaccination failure灭活苗免疫失败Challenge test-verify protection 攻毒试验验证保护力Take samples at a later moment过一段时间再次采样第二十五页，讲稿共七十三页哦Interpretation:qualitative tests分析：定性试验NegativoPositive阳性Sample阳性Infection感染Sero-response to the antigen 对抗原血清学反应Confirmation确诊Use a test with a higher specificity 使用敏感性更高的试验Isolation of the agent病原分离Vaccine疫苗live field challenge or vaccine response活毒野毒感染或疫苗反应killed vaccine response灭活苗疫苗反应第二十六页，讲稿共七十三页哦Interpretation:quantitative tests分析：定量试验NegativoHigh titre高滴度Sample样品Vaccine疫苗live field challenge or vaccine response活毒野毒感染或疫苗反应killed vaccine response灭活苗疫苗反应Confirmation确诊Use a test with a higher specificity 使用敏感性更高的试验Isolation of the agent病原分离Infection感染Sero-response to the antigen 对抗原血清学反应第二十七页，讲稿共七十三页哦Vaccine疫苗Good response to the vaccinationGood vaccine coverage接种后的良好反应Interpretation:(semi-)quantitative tests分析：半定量试验Low CV低CVNarrow distribution of titres 狭窄的滴度分布No infection没有感染Chronic Infection in the population鸡群内慢性感染Samples第二十八页，讲稿共七十三页哦Vaccine疫苗poor response to the vaccination.接种免疫反应差，免疫覆盖差Poor vaccination coverageInterpretation:(semi-)quantitative tests分析：半定量试验Wide CVCV值大A very wide range of titres滴度分布范围非常宽Acute Infection急性感染A chronic infection in some indivduals某些个体慢性感染samplesCheck the batch检查批号Compare with field results与临床结果比较第二十九页，讲稿共七十三页哦Interpretation in broilers对肉鸡群的分析Levels of Maternally Derived Antibodies 母源抗体的水平Knowledge about the sanitary statusOutbreaks.Infections or challenge 关于卫生状况的知识 疾病爆发、感染、攻毒Check the vaccination programmesFixed dates检查免疫程序固定日期第三十页，讲稿共七十三页哦Normal Curve正态曲线第三十一页，讲稿共七十三页哦Comparing curves曲线比较第三十二页，讲稿共七十三页哦Interpretation chickens or live vaccines分析雏鸡或活苗血清学结果外表性能分析措施第三十三页，讲稿共七十三页哦Low titre+CV good+good performance低滴度良好的CV良好的性能=continue with the control programme继续控制程序第三十四页，讲稿共七十三页哦High Titre+good performance高滴度良好的性能=continue the control programme andKeep an eye on the biosecurity!继续控制程序，注意生物安全！Watch out!注意Vaccinated flock免疫过的鸡群=Infection with protection 具保护力的感染=Vaccination with a hot strain 用强毒接种Non-vaccinated flock未免疫的鸡群=past infection 过去感染过=Infection with a mild strain 弱毒株感染过第三十五页，讲稿共七十三页哦High titre+narrow CV高滴度窄CVOld infectionGood vaccine coveragewith the use of a strong vaccine strain使用了强的疫苗毒株，具有良好的疫苗覆盖性第三十六页，讲稿共七十三页哦High titre+wide CV高滴度宽CVAcute infection急性感染Poor vaccine coverage using a hot vaccine strain 使用了强毒疫苗，疫苗覆盖性差。第三十七页，讲稿共七十三页哦High titre+poor performance高滴度差性能Isolate and identify the agent.分离鉴定病毒Improve the control programme.改进控制程序Improve the biosecurity改进生物安全。=fieldinfection Challenge and disease野毒感染和疾病第三十八页，讲稿共七十三页哦Low titre+poor performance低滴度差性能 acute infection no antibodies yet 急性感染还未产生抗体 other problems其它问题Retest 1 week later 1周后重新检测Look for other possible cause 寻找其它可能的问题第三十九页，讲稿共七十三页哦Interpretation by Agent按病原分析第四十页，讲稿共七十三页哦Interpretation in breeders or layers种鸡和蛋鸡的分析Knowledge of the sanitary statusOutbreaks,infection or challenge?卫生状况的知识爆发、感染、攻毒Check vaccination programmesPre-fixed dates检查免疫程序Comparison with production curves 比较生产曲线第四十一页，讲稿共七十三页哦High and low titres-how much?第四十二页，讲稿共七十三页哦It is all is a matter of perspective第四十三页，讲稿共七十三页哦High or low titres-how high?How low?抗体滴度高和低的认识Base lines基准线第四十四页，讲稿共七十三页哦High or low titres-cuanto?滴度高或低多少？Base de datos数据的基准线第四十五页，讲稿共七十三页哦Curves-IBD(Real)IBD的抗体滴度曲线第四十六页，讲稿共七十三页哦Serology of the individual 个体血清学Serology of populations(flocks)群体血清学Samples采样数量8Baseline-minimum 23,ideal 28 基准线最低23份，理想28份 8Monitoring-minimum 15 监测最低15份8Diagnostics-between 28 and 146 诊断（28146）份第四十七页，讲稿共七十三页哦Serum“banks”血清保存Research研究Pruebas comprobatoriasImportant for diagnostics诊断的重要性Security安全性Adequate storage of the serum avoid evaporation 正确存储血清，避免蒸发第四十八页，讲稿共七十三页哦Collecting samples采集 样品Monitoring监测Consistence 时间安排Vaccination Programmes 免疫程序Mineral-oil vaccines 矿物油疫苗Feed 饲料Diagnostics诊断Paired samples双份采样SerotecaConditions under which the samples are taken-false positives 样品采集的环境条件（可能造成假阳性）Conditions under which the samples have been taken and the storage第四十九页，讲稿共七十三页哦ELISA titresELISA滴度A lot of variation depends on the author and the kit 根据不同的操作人员和试剂盒，ELISA滴度会有差异第五十页，讲稿共七十三页哦ELISA-IDEXX第五十一页，讲稿共七十三页哦ELISA-KPL第五十二页，讲稿共七十三页哦ELISA-IDEXX第五十三页，讲稿共七十三页哦ELISA-KPL第五十四页，讲稿共七十三页哦ELISA-IDEXX第五十五页，讲稿共七十三页哦ELISA-KPL第五十六页，讲稿共七十三页哦Conclusion(any questions?)第五十七页，讲稿共七十三页哦第五十八页，讲稿共七十三页哦The End第五十九页，讲稿共七十三页哦Ay ay ay ayeven more veterinarians!第六十页，讲稿共七十三页哦第六十一页，讲稿共七十三页哦Curvas 第六十二页，讲稿共七十三页哦Interpretatin of resultsGMTCV第六十三页，讲稿共七十三页哦Mathematical problems in the calculation of the coefficient of variation For formula reasons low mean valueswill result in high CV valuesJudge the histogramme第六十四页，讲稿共七十三页哦Distribution of titresGMT-igualesCV-narrowCV wideCV-wide第六十五页，讲稿共七十三页哦第六十六页，讲稿共七十三页哦第六十七页，讲稿共七十三页哦第六十八页，讲稿共七十三页哦第六十九页，讲稿共七十三页哦Curves birds in Production第七十页，讲稿共七十三页哦Curve-Production第七十一页，讲稿共七十三页哦Curve-Production(2)第七十二页，讲稿共七十三页哦感谢大家观看第七十三页，讲稿共七十三页哦