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开发一种基于单克隆抗体的竞争酶联免疫吸附测定法,用于检测和滴定小反刍兽疫(PPR)病毒抗体。

Development of a monoclonal antibody based competitive-ELISA for detection and titration of antibodies to peste des petits ruminants (PPR) virus.

作者信息

Singh R P, Sreenivasa B P, Dhar P, Shah L C, Bandyopadhyay S K

机构信息

Division of Virology, Indian Veterinary Research Institute, Mukteswar, Nainital 263 138, Uttaranchal, India.

出版信息

Vet Microbiol. 2004 Jan 14;98(1):3-15. doi: 10.1016/j.vetmic.2003.07.007.

Abstract

Peste des petits ruminants (PPR) is an acute febrile, viral, disease of small ruminants with great economic importance. A competitive-ELISA (c-ELISA) test was developed for detection of antibodies to PPR virus in the sera samples of goats and sheep. The test uses monoclonal antibody to a neutralizing epitope of haemagglutinin protein of the virus. Based on the distribution of known negative sera samples (n=933) in respect of PPR virus antibodies in the test, a cut-off value was set as 38%. This value was the result of mean of negative population added with two times the standard deviations. A total of 1668 sera samples from goat and sheep and 32 sera from cattle were screened by c-ELISA and virus neutralization test (VNT). Efficacy of c-ELISA compared very well with VNT having high relative specificity (98.4%) and sensitivity (92.4%). The sensitivity of c-ELISA for PPR sero-surveillance could further be increased (95.4%), if the target population is non-vaccinated. c-ELISA test correlated well with VNT (r=0.845) for end-point titration of PPR virus antibody in 64 goat sera samples. It could clearly separate infected population from uninfected in field sera. Using c-ELISA test paired sera samples from 13 goats provided a clear diagnosis of PPR virus infection. Furthermore, antibodies to PPR virus could be successfully detected during 1 year after vaccination in four goats inoculated with an experimental PPR vaccine. Findings suggest that the c-ELISA test developed can easily replace VNT for sero-surveillance, sero-monitoring, diagnosis from paired sera samples and end-point titration of PPR virus antibodies.

摘要

小反刍兽疫(PPR)是一种对小反刍动物具有高度经济重要性的急性发热性病毒性疾病。开发了一种竞争ELISA(c-ELISA)检测方法,用于检测山羊和绵羊血清样本中抗PPR病毒的抗体。该检测方法使用针对该病毒血凝素蛋白中和表位的单克隆抗体。根据检测中已知的933份PPR病毒抗体阴性血清样本的分布情况,将临界值设定为38%。该值是阴性群体平均值加上两倍标准差的结果。通过c-ELISA和病毒中和试验(VNT)对1668份山羊和绵羊血清样本以及32份牛血清进行了筛查。c-ELISA与VNT的效果相比非常好,具有较高的相对特异性(98.4%)和敏感性(92.4%)。如果目标群体未接种疫苗,c-ELISA对PPR血清监测的敏感性可进一步提高(95.4%)。在64份山羊血清样本中,c-ELISA检测与VNT在PPR病毒抗体终点滴定方面相关性良好(r=0.845)。它可以在现场血清中清晰地将感染群体与未感染群体区分开来。使用c-ELISA检测,来自13只山羊的配对血清样本对PPR病毒感染做出了明确诊断。此外,在接种实验性PPR疫苗的4只山羊中,接种后1年内成功检测到了抗PPR病毒抗体。研究结果表明,所开发的c-ELISA检测方法可轻松替代VNT进行血清监测、血清监测、配对血清样本诊断以及PPR病毒抗体终点滴定。

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