Hayatullah Khan , Farhan Anwar Khan , Umer Sadique , Ijaz Ahmad , Muhammad Saeed , Faisal Ahmad , Faiz Ur Rehman , Hanif Ur Rahman
{"title":"开发和验证用于山羊传染性绵羊胸膜肺炎血清诊断的间接酶联免疫吸附测定法","authors":"Hayatullah Khan , Farhan Anwar Khan , Umer Sadique , Ijaz Ahmad , Muhammad Saeed , Faisal Ahmad , Faiz Ur Rehman , Hanif Ur Rahman","doi":"10.1016/j.smallrumres.2024.107271","DOIUrl":null,"url":null,"abstract":"<div><p><em>Mycoplasma capricolum</em> subsp. <em>capripneumoniae</em> (Mccp) is a dreadful pathogen of contagious caprine pleuropneumonia (CCPP). Mccp local strains have recently been isolated from naturally infected goats in northern areas of Pakistan. In this study, we established an in-house indirect ELISA (iELISA) based on whole-cell proteins (WCP) of local Mccp strains for the detection of anti-Mccp antibodies. Serum samples from naturally (n=500) and experimentally infected (n=20) and non-infected animals (n=100) were used for the establishment of iELISA. The specificity and sensitivity of the iELISA were determined using a commercially available competitive ELISA (cELISA) kit as a reference standard. The cut-off value (OD<sub>450</sub>) was found 0.53 by ROC analysis. As indicated by a kappa value of 0.88 and 96.66% agreement, both assays showed perfect agreement. An iELISA based on WCP of local Mccp detected seroconversion in experimental goats 7 days post-infection (dpi). The iELISA revealed a sensitivity of 90.83% (95% CI: 83.77%, 95.55%) and a specificity of 92.31% (95% CI: 84.79%, 96.85%) and has no cross-reaction with related mycoplasma epidemic diseases. For both inter-assay and intra-assay precision, the iELISA percentage coefficient of variation (%CV) was 4.8% and 1.74%, respectively. In conclusion, this in-house iELISA based on local Mccp-WCP was determined as a sensitive and specific assay for the diagnosis of Mccp infection.</p></div>","PeriodicalId":21758,"journal":{"name":"Small Ruminant Research","volume":null,"pages":null},"PeriodicalIF":1.6000,"publicationDate":"2024-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development and validation of an indirect ELISA for the serodiagnosis of contagious caprine pleuropneumonia in goats\",\"authors\":\"Hayatullah Khan , Farhan Anwar Khan , Umer Sadique , Ijaz Ahmad , Muhammad Saeed , Faisal Ahmad , Faiz Ur Rehman , Hanif Ur Rahman\",\"doi\":\"10.1016/j.smallrumres.2024.107271\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><em>Mycoplasma capricolum</em> subsp. <em>capripneumoniae</em> (Mccp) is a dreadful pathogen of contagious caprine pleuropneumonia (CCPP). Mccp local strains have recently been isolated from naturally infected goats in northern areas of Pakistan. In this study, we established an in-house indirect ELISA (iELISA) based on whole-cell proteins (WCP) of local Mccp strains for the detection of anti-Mccp antibodies. Serum samples from naturally (n=500) and experimentally infected (n=20) and non-infected animals (n=100) were used for the establishment of iELISA. The specificity and sensitivity of the iELISA were determined using a commercially available competitive ELISA (cELISA) kit as a reference standard. The cut-off value (OD<sub>450</sub>) was found 0.53 by ROC analysis. As indicated by a kappa value of 0.88 and 96.66% agreement, both assays showed perfect agreement. An iELISA based on WCP of local Mccp detected seroconversion in experimental goats 7 days post-infection (dpi). The iELISA revealed a sensitivity of 90.83% (95% CI: 83.77%, 95.55%) and a specificity of 92.31% (95% CI: 84.79%, 96.85%) and has no cross-reaction with related mycoplasma epidemic diseases. For both inter-assay and intra-assay precision, the iELISA percentage coefficient of variation (%CV) was 4.8% and 1.74%, respectively. In conclusion, this in-house iELISA based on local Mccp-WCP was determined as a sensitive and specific assay for the diagnosis of Mccp infection.</p></div>\",\"PeriodicalId\":21758,\"journal\":{\"name\":\"Small Ruminant Research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.6000,\"publicationDate\":\"2024-04-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Small Ruminant Research\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0921448824000774\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"AGRICULTURE, DAIRY & ANIMAL SCIENCE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Small Ruminant Research","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0921448824000774","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
Development and validation of an indirect ELISA for the serodiagnosis of contagious caprine pleuropneumonia in goats
Mycoplasma capricolum subsp. capripneumoniae (Mccp) is a dreadful pathogen of contagious caprine pleuropneumonia (CCPP). Mccp local strains have recently been isolated from naturally infected goats in northern areas of Pakistan. In this study, we established an in-house indirect ELISA (iELISA) based on whole-cell proteins (WCP) of local Mccp strains for the detection of anti-Mccp antibodies. Serum samples from naturally (n=500) and experimentally infected (n=20) and non-infected animals (n=100) were used for the establishment of iELISA. The specificity and sensitivity of the iELISA were determined using a commercially available competitive ELISA (cELISA) kit as a reference standard. The cut-off value (OD450) was found 0.53 by ROC analysis. As indicated by a kappa value of 0.88 and 96.66% agreement, both assays showed perfect agreement. An iELISA based on WCP of local Mccp detected seroconversion in experimental goats 7 days post-infection (dpi). The iELISA revealed a sensitivity of 90.83% (95% CI: 83.77%, 95.55%) and a specificity of 92.31% (95% CI: 84.79%, 96.85%) and has no cross-reaction with related mycoplasma epidemic diseases. For both inter-assay and intra-assay precision, the iELISA percentage coefficient of variation (%CV) was 4.8% and 1.74%, respectively. In conclusion, this in-house iELISA based on local Mccp-WCP was determined as a sensitive and specific assay for the diagnosis of Mccp infection.
期刊介绍:
Small Ruminant Research publishes original, basic and applied research articles, technical notes, and review articles on research relating to goats, sheep, deer, the New World camelids llama, alpaca, vicuna and guanaco, and the Old World camels.
Topics covered include nutrition, physiology, anatomy, genetics, microbiology, ethology, product technology, socio-economics, management, sustainability and environment, veterinary medicine and husbandry engineering.