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荧光偏振检测法与竞争 ELISA 诊断牛布鲁氏菌病的比较评估及其血清监测。

Comparative evaluation of fluorescence polarization assay and competitive ELISA for the diagnosis of bovine brucellosis vis-a-vis sero-monitoring.

机构信息

Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR- NIVEDI), Yelahanka, Bengaluru 560 064, India; Jain University, Centre for Post Graduate Studies, Jayanagar, Bengaluru 560041; India.

Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR- NIVEDI), Yelahanka, Bengaluru 560 064, India.

出版信息

J Microbiol Methods. 2020 Mar;170:105858. doi: 10.1016/j.mimet.2020.105858. Epub 2020 Jan 31.

Abstract

Brucellosis is an important zoonosis that constitutes a serious public health hazard which is caused by a bacterium belonging to the genus Brucella. In the present study, two highly specific serological tests for brucellosis diagnosis, fluorescence polarization assay (FPA) and competitive ELISA (cELISA) were standardized in the laboratory, evaluated and compared with rose bengal plate test (RBPT), indirect ELISA (iELISA) and commercial cELISA kit. For test evaluation, 1386 serum samples [apparently healthy animals (n = 260), samples from Brucella infected farms (n = 701) and B. abortus S19 vaccinated animals (n = 425)] were analyzed to assess suitable diagnostic test in B. abortus S19 post vaccinated bovine population. In apparently healthy brucellosis free farms, RBPT, iELISA, in-house FPA and cELISA were found to be highly specific than commercial cELISA. Commercial cELISA kit was comparatively more sensitive than other serological tests in samples collected from infected farms. The FPA showed sensitivity nearly equal to RBPT and in-house cELISA showed greater sensitivity than RBPT in infected farms. In animals with persistent vaccinal antibodies, only in-house FPA and cELISA recorded higher specificity of 87.64 and 90.27%, respectively. The other tests, RBPT and iELISA displayed similar reactivity with vaccine antibodies to that of infection antibodies whereas commercial cELISA kit showed an intermediate specificity of 47.69%. With these findings, RBPT, iELISA and cELISA are suggested for screening infected herds, and in-house developed FPA and cELISA tests with a proven specificity can be used for confirmatory diagnosis of brucellosis in B. abortus S19 post vaccinated animal populations.

摘要

布鲁氏菌病是一种重要的人畜共患病,由布鲁氏菌属细菌引起,对公共卫生构成严重威胁。本研究对两种用于布鲁氏菌病诊断的高特异性血清学检测方法——荧光偏振检测(FPA)和竞争酶联免疫吸附试验(cELISA)进行了实验室标准化、评估,并与虎红平板凝集试验(RBPT)、间接酶联免疫吸附试验(iELISA)和商业 cELISA 试剂盒进行了比较。为了进行检测评估,对 1386 份血清样本[表现健康的动物(n=260)、布鲁氏菌感染场的样本(n=701)和 B.abortus S19 疫苗接种动物(n=425)]进行了分析,以评估 B.abortus S19 疫苗接种牛群中适合的诊断检测方法。在无明显布鲁氏菌病的健康农场中,RBPT、iELISA、内部 FPA 和 cELISA 的特异性均高于商业 cELISA。在感染场采集的样本中,商业 cELISA 试剂盒的敏感性比其他血清学检测方法都高。FPA 的敏感性与 RBPT 相近,而在感染场中,内部 cELISA 的敏感性高于 RBPT。在持续存在疫苗抗体的动物中,只有内部 FPA 和 cELISA 记录的特异性分别为 87.64%和 90.27%较高。其他检测方法,如 RBPT 和 iELISA,与感染抗体的反应性与疫苗抗体相似,而商业 cELISA 试剂盒的特异性为 47.69%。根据这些发现,建议 RBPT、iELISA 和 cELISA 用于筛选感染牛群,而内部开发的具有特异性的 FPA 和 cELISA 检测方法可用于 B.abortus S19 疫苗接种动物群体中布鲁氏菌病的确认诊断。

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