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

IF 1.6 3区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Small Ruminant Research Pub Date : 2024-04-17 DOI:10.1016/j.smallrumres.2024.107271
Hayatullah Khan , Farhan Anwar Khan , Umer Sadique , Ijaz Ahmad , Muhammad Saeed , Faisal Ahmad , Faiz Ur Rehman , Hanif Ur Rahman
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引用次数: 0

摘要

冠状支原体冠状病毒亚种(Mccp)是传染性山羊胸膜肺炎(CCPP)的一种可怕病原体。最近,从巴基斯坦北部地区自然感染的山羊中分离出了 Mccp 地方菌株。在这项研究中,我们建立了一种基于当地 Mccp 株系全细胞蛋白(WCP)的内部间接 ELISA(iELISA)方法,用于检测抗 Mccp 抗体。iELISA的建立采用了自然感染(n=500)、实验感染(n=20)和非感染(n=100)动物的血清样本。使用市售竞争性 ELISA(cELISA)试剂盒作为参考标准,测定 iELISA 的特异性和灵敏度。通过 ROC 分析发现,临界值(OD450)为 0.53。卡帕值为 0.88,吻合率为 96.66%,表明两种检测方法完全吻合。基于本地 Mccp WCP 的 iELISA 检测出了感染后 7 天(dpi)实验山羊的血清转换。iELISA 的灵敏度为 90.83%(95% CI:83.77%,95.55%),特异度为 92.31%(95% CI:84.79%,96.85%),与相关支原体流行病无交叉反应。在测定间和测定内精密度方面,iELISA 的百分比变异系数(%CV)分别为 4.8%和 1.74%。总之,这种基于本地 Mccp-WCP 的内部 iELISA 被确定为诊断 Mccp 感染的灵敏而特异的检测方法。
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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.

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来源期刊
Small Ruminant Research
Small Ruminant Research 农林科学-奶制品与动物科学
CiteScore
3.10
自引率
11.10%
发文量
210
审稿时长
12.5 weeks
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.
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