试验室检测及数据分析应用.ppt

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Interpretation of Serological results,血清结果的分析,Dra.Beatriz Cardoso,MSc,ACPV,Dr.Oscar Morales,ACPV,Lohmann Animal Health,德国罗曼动物保健品有限公司,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?,broilers,Layers 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,What is serology?,什么是血清学？,Detection of antibodies,抗体的检测,Types of antibodies,抗体的类型,IgA,IgM,IgE,IgG,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,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 2,x,10,x(2,log,10,log),结果表达为,2,x,10,x(2,log,10,log),HI,Serum neutralization,HI,血清中和试验,S,pecificity,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,Ideal,理想状况,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 100,Sensibility=98/(98+2)x100=98%,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 100,specificity=80/(80+20)x100=80%,Calculations,计算,Sensibility=98%,specificity=80%,敏感性,98,特异性,80,Aplication in the field,现场运用,Farm with 10.000 birds,某场,10000,羽鸡,100%checked,100,进行检测,500 positives,出现,500,阳性,490,true positives,10,false,positives,490,真阳性,10,假阳性,specific test,特异性试验,Calculations 2,计算,2,Sensibility=80/(100)x100=80%,敏感性,80/(100)x100=80%,specificity=100/(100)x 100=100%,特异性,100/(100)x 100=100%,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,Test commonly used in poultry,家禽常用的检测方法,RPA-Mycoplasma,Salmonella,AGP-IBV,IBD,Marek,Reo,HI-,ND,IBV,EDS,Mycoplasma,Coryza,Test commonly used in poultry,家禽常用的检测方法,SN-IBV,IBD,all for research purposes,ELISA nearly all agents,Serology,血清学,Diagnostics,诊断,Monitoring,监测,INTERPRETATION,分析,Criteria for interpretation,分析的标准,What disease do I want to verify?,我要验证什么病？,For what purpose?Diagnostics or Monitoring?,分析的目的是什么？诊断或监测？,What type of exploitation do I have?,我该采取什么方法？,How should I interpret the results,我该怎样分析结果？,Dogma o Paradigma?,USUALLY THE HIGHER THE TITRE,THE BETTER THE PROTECTION,通常滴度越高，保护力越高,Basis for the interpretation,分析的基础,Presence of antibodies,Infection,Development of the immunity,存在抗体,感染,免疫力的产生,Absence of antibodies,There is no infection,The Infection was very recent,The 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,Interpretation:qualitative tests,分析：定性试验,Negative,阴性,Positivo,S,ample,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,V,accine,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,Interpretation:(semi-)quantitative tests,分析：半定量试验,Negative or low titre,阴性或低滴度,Positivo,S,ample,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,V,accine,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,Interpretation:qualitative tests,分析：定性试验,Negativo,Positive,阳性,S,ample,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,V,accine,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,Interpretation:quantitative tests,分析：定量试验,Negativo,High titre,高滴度,S,ample,样品,V,accine,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,V,accine,疫苗,Good,response,to the vaccination,Good vaccine,coverage,接种后的良好反应,Interpretation:(semi-)quantitative tests,分析：半定量试验,Low CV,低,CV,Narrow 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,S,ample,s,Vaccine,疫苗,poor,response,to the vaccination,.,接种免疫反应差，免疫覆盖差,Poor vaccination coverage,Interpretation:(semi-)quantitative tests,分析：半定量试验,Wide CV,CV,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,samples,Check the,batch,检查批号,Compare,with field,results,与临床结果比较,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 status,Outbreaks.Infections or challenge,关于卫生状况的知识,疾病爆发、感染、攻毒,Check the vaccination programmes,Fixed dates,检查免疫程序,固定日期,Normal Curve,正态曲线,Comparing curves,曲线比较,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,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,High Titre+good performance,高滴度良好的性能,=continue the control programme and,Keep 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,High titre+narrow CV,高滴度窄,CV,Old infection,Good vaccine,coverage,with 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,High titre+wide CV,高滴度宽,CV,Acute,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。,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,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,Interpretation by Agent,按病原分析,Interpretation in breeders or layers,种鸡和蛋鸡的分析,Knowledge of the sanitary status,Outbreaks,infection or challenge?,卫生状况的知识,爆发、感染、攻毒,Check vaccination programmes,Pre-fixed dates,检查免疫程序,Comparison with production curves,比较生产曲线,High and low titres-how much?,It is all is a matter of perspective,High or low titres-how high?How low?,抗体滴度高和低的认识,Base lines,基准线,High or low titres-cuanto?,滴度高或低多少？,Base de datos,数据的基准线,Curves-IBD(Real),IBD,的抗体滴度曲线,Serology of the,individual,个体血清学,Serology of populations,(,flocks,),群体血清学,Samples,采样数量,Baseline,-,minimum,23,ideal 28,基准线最低,23,份，理想,28,份,Monitoring,-,minimum,15,监测最低,15,份,Diagnostics,-,between,28,and,146,诊断（,28,146,）份,Serum“banks”,血清保存,R,esearch,研究,Pruebas comprobatorias,Important 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,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,F,eed,饲料,Diagnostics,诊断,Paired samples,双份采样,Seroteca,Conditions 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,ELISA titres,ELISA,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,ELISA-IDEXX,ELISA-KPL,ELISA-IDEXX,ELISA-KPL,ELISA-IDEXX,ELISA-KPL,Conclusion,(any questions?),Thank you,The End,Ay ay ay ay,even more veterinarians!,Curvas,Interpretatin of results,GMT,CV,Mathematical problems in the calculation of the coefficient of variation,For formula reasons low mean values,will result in high CV values,Judge the histogramme,Distribution of titres,GMT-iguales,CV-,narrow,CV,wide,CV-,wide,Curves birds in Production,Curve-Production,Curve-Production(2),