采用粪便培养、乳汁培养和粪便聚合酶链反应对印度高灵敏度本土牛奶酶联免疫吸附测定试剂盒在牛副结核病（BJD）诊断中的评估。

Evaluation of highly sensitive indigenous milk ELISA kit with fecal culture, milk culture and fecal-PCR for the diagnosis of bovine Johne's disease (BJD) in India.

作者信息

Singh S V, Singh A V, Singh R, Sandhu K S, Singh P K, Sohal J S, Gupta V K, Vihan V S

机构信息

Goat Health Division, Central Institute for Research on Goats, Makhdoom, P.O. Farah, Mathura, UP 281 122, India.

出版信息

Comp Immunol Microbiol Infect Dis. 2007 May;30(3):175-86. doi: 10.1016/j.cimid.2006.12.002. Epub 2007 Feb 27.

DOI:10.1016/j.cimid.2006.12.002

PMID:17328952

Abstract

Country lacks indigenous diagnostic kits against Johne's disease in animals. Indigenous ELISA and IS 900 PCR kits, originally developed for goats and sheep, have been adapted for screening of lactating cows. Multiple diagnostic tests were used to screen 26 lactating dairy cows against Johne's disease. Milk ELISA was evaluated with fecal culture, milk culture and fecal PCR. Of the 26 samples from lactating cows, 84.6, 96.1, 88.4 and 23.0% were positive in fecal culture, milk culture, m-ELISA and m-PCR, respectively. Comparatively milk sediment and milk fat culture detected 84.6 and 76.9% cows positive, respectively. Comparatively fecal culture and milk culture detected 84.6 and 96.1% cows positive, respectively. M-ELISA detected 11.5, 0.0, 11.5, 61.0 and 15.3%, cows as negative, suspected, low positive, positive and strong positive, respectively. There was good correlation between milk and fecal culture with m-ELISA. Three negative cows in m-ELISA were also detected in milk and fecal culture. Of the 26 decontaminated fecal samples, 23.0% cows were positive using specific IS 900 f-PCR. Comparative evaluation of m-ELISA with fecal and milk culture showed agreement in 80.7 and 84.6% cows, respectively. Sensitivity of m-ELISA with respect to fecal and culture was 90.9 and 95.6%, respectively. Comparative evaluation of four tests (milk culture, fecal culture, m-ELISA and f-PCR) showed that only 15.3% cows were detected in all the four tests. In three tests (fecal and milk culture and m-ELISA), 57.6% cows were detected positive. None of the cow was exclusively detected in f-PCR. Of the four diagnostic tests used milk culture was most sensitive (96.15%), followed by fecal culture (86.6%), m-ELISA (76.9%) and IS 900 PCR (23.0%) for the diagnosis of bovine Johne's disease (BJD). Milk ELISA detected only one cow extra, which was negative in milk culture. In view of the simplicity, rapidity and efficacy present milk ELISA kit employing soluble protoplasmic antigen from native Map 'Bison type' genotype of goat origin can be reliable for screening of bovine population against Johne's disease in India.

摘要

该国缺乏针对动物约翰氏病的本土诊断试剂盒。最初为山羊和绵羊开发的本土酶联免疫吸附测定（ELISA）和IS 900聚合酶链反应（PCR）试剂盒已被用于筛选泌乳奶牛。使用多种诊断测试对26头泌乳奶牛进行约翰氏病筛查。用粪便培养、乳汁培养和粪便PCR对乳汁ELISA进行评估。在26份泌乳奶牛样本中，粪便培养、乳汁培养、乳汁ELISA和乳汁PCR的阳性率分别为84.6%、96.1%、88.4%和23.0%。相比之下，乳汁沉淀物和乳脂培养检测出的阳性奶牛分别为84.6%和76.9%。相比之下，粪便培养和乳汁培养检测出的阳性奶牛分别为84.6%和96.1%。乳汁ELISA检测出的奶牛阴性、疑似、低阳性、阳性和强阳性比例分别为11.5%、0.0%、11.5%、61.0%和15.3%。乳汁培养和粪便培养与乳汁ELISA之间存在良好的相关性。乳汁ELISA检测为阴性的3头奶牛在乳汁和粪便培养中也被检测出来。在26份经过净化的粪便样本中，使用特异性IS 900荧光定量PCR检测出23.0%的奶牛呈阳性。乳汁ELISA与粪便和乳汁培养的比较评估显示，分别有80.7%和84.6%的奶牛结果一致。乳汁ELISA相对于粪便培养和乳汁培养的灵敏度分别为90.9%和95.6%。对四项检测（乳汁培养、粪便培养、乳汁ELISA和荧光定量PCR）的比较评估显示，所有四项检测均检测出阳性的奶牛仅为15.3%。在三项检测（粪便和乳汁培养以及乳汁ELISA）中，57.6%的奶牛检测为阳性。荧光定量PCR未单独检测出任何一头奶牛呈阳性。在所使用的四项诊断测试中，乳汁培养对牛约翰氏病（BJD）的诊断最为敏感（96.15%），其次是粪便培养（86.6%）、乳汁ELISA（76.9%）和IS 900 PCR（23.0%）。乳汁ELISA仅多检测出一头奶牛呈阳性，该奶牛在乳汁培养中为阴性。鉴于其简便性、快速性和有效性，目前采用源自山羊“野牛型”基因型天然副结核分枝杆菌可溶性原生质体抗原的乳汁ELISA试剂盒，对于印度牛群约翰氏病的筛查可能是可靠的。