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应用种特异性 ELISA 对感染奶牛群中地方性柯克斯体感染的动态分析。

Insights into the dynamics of endemic Coxiella burnetii infection in cattle by application of phase-specific ELISAs in an infected dairy herd.

机构信息

Bavarian Animal Health Service, Senator-Gerauer-Straße 23, D-85586 Poing, Bavaria, Germany.

出版信息

Vet Microbiol. 2011 Aug 5;151(3-4):291-300. doi: 10.1016/j.vetmic.2011.03.007. Epub 2011 Mar 16.

Abstract

Serological diagnosis of acute and chronic Q fever in humans relies on detection of antibodies to phase I (PhI) and II (PhII) antigens of Coxiella (C.) burnetii. Although phase-specific antigens are available, they are not yet used in ruminants as they are in humans. This study focuses on phase-specific serology as a tool for analysis of the dynamics of infection in cattle. As a prerequisite, sero-prevalence in Bavarian cattle (1) and sero-prevalences for age-groups (2) were determined by ELISA (CHEKIT Q-Fever; mix of PhI/PhII-antigen). Subsequently, phase-specific antigens were coated onto ELISA plates individually and tests were simultaneously applied in an endemically infected herd with about 90 dairy cows and 250 calves/heifers in April 2005, March 2006 and retrospectively in May and October 2004. From April 2005 onward, placentas were analysed for C. burnetii by PCR (3). (1) Sero- and herd prevalences based on 21,051 sera from 603 Bavarian dairy farms collected in 2003 were 14.8% ± 0.48% and 72.3% ± 3.6%, respectively. (2) Analysis of 3965 animals from 105 farms for which age was reported revealed a base level of sero-prevalence of less than 5% in 1-2 years old animals, it increased to 15% in 2-3 years old and reached a plateau (25-30%) in cows four years and older. (3) In May 2004 and April 2005 a peak of PhI(-)/PhII(+)-prevalence in primiparous cows (2.0-3.5 years) was observed; but not in October 2004 and March 2006. The PhI(-)/PhII(+)-pattern in primiparous cows changed to negative (one-third), PhI(+)/PhII(+) (1/3) or persisted (1/3). In contrast, sero-conversion was rare in multiparous cows (>3.5 years). If the PhI(-)/PhII(+) pattern was detected, it was due to an infection in preceding years. This pattern persisted (2/3) or changed to negative (1/3); a change to PhI(+)/PhII(+) did not occur. PhI(-)/PhII(+) in heifers (1-2 years) always changed to negative. Detection of PhII-antibodies was significantly associated with PCR-positive placentas. Remarkably, 45% of sera with the PhI(-)/PhII(+) pattern were negative for the CHEKIT Q-Fever ELISA, thus this test missed an important group of infected animals.

摘要

人感染急性和慢性 Q 热的血清学诊断依赖于对贝纳柯克斯体(Coxiella)I 相(PhI)和 II 相(PhII)抗原的抗体检测。虽然已经有了针对各相的特异性抗原,但在反刍动物中尚未像在人类中那样使用。本研究主要关注各相特异性血清学作为分析牛群感染动态的工具。作为前提条件,通过 ELISA(CHEKIT Q-Fever;PhI/PhII 抗原混合物)确定了巴伐利亚牛的血清流行率(1)和年龄组的血清流行率(2)。随后,单独将各相特异性抗原包被到 ELISA 板上,并于 2005 年 4 月、2006 年 3 月和 2004 年 5 月和 10 月在一个地方性感染的牛群中同时进行了检测,该牛群约有 90 头奶牛和 250 头犊牛/小母牛。自 2005 年 4 月起,通过 PCR(3)对胎盘进行了贝纳柯克斯体分析。(1)基于 2003 年从 603 个巴伐利亚奶牛场收集的 21051 份血清进行的血清和群体流行率分别为 14.8%±0.48%和 72.3%±3.6%。(2)对 105 个有年龄报告的农场的 3965 头动物进行分析,结果表明 1-2 岁动物的基础血清流行率低于 5%,2-3 岁动物的血清流行率增加到 15%,4 岁及以上的牛达到了 25-30%的平台期。(3)2004 年 5 月和 2005 年 4 月,初产奶牛(2-3 岁)中观察到 PhI(-)/PhII(+)流行率的高峰(2.0-3.5 岁);但 2004 年 10 月和 2006 年 3 月没有观察到。初产奶牛的 PhI(-)/PhII(+)模式转为阴性(三分之一)、PhI(+)/PhII(+)(三分之一)或持续存在(三分之一)。相比之下,多产奶牛(>3.5 岁)的血清转化率很少。如果检测到 PhI(-)/PhII(+)模式,则是由于前几年的感染。该模式持续存在(三分之二)或转为阴性(三分之一);没有发生 PhI(+)/PhII(+)的转变。1-2 岁小母牛的 PhI(-)/PhII(+)模式始终转为阴性。检测到 PhII 抗体与 PCR 阳性胎盘显著相关。值得注意的是,45%的 PhI(-)/PhII(+)模式血清对 CHEKIT Q-Fever ELISA 呈阴性,因此该检测漏掉了一个重要的感染动物群体。

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