Evaluation of genus-specific and species-specific real-time PCR assays for the identification of Brucella spp.
Evaluation of genus-specific and species-specific real-time PCR assays for the identification of Brucella spp.
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DOI：
10.1515/cclm.2007.305
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发表时间：
2007-01-01
影响因子：
6.8
通讯作者：
Tomaso, Herbert
中科院分区：
文献类型：
--
作者：
Al Dahouk, Sascha;Noeckler, Karsten;Tomaso, Herbert
背景：使用常规微生物学技术鉴定布鲁氏菌分离株是耗时且危险的。因此，我们评估了实时PCR检测在种属水平上鉴定布鲁氏菌属成员的性能。方法：我们评估了一种内部开发的检测方法和多种先前发表的靶向bcsp 31、per、IS 711的实时PCR检测方法，alkB/IS 711和BMEI 1162/IS 711使用代表所有物种的生物型的248株布鲁氏菌菌株和一大组临床相关、遗传学相关和血清学交叉反应细菌。未观察到错误识别。然而，一些已发表的检测方法未检出牛种布鲁氏菌和猪种布鲁氏菌的几种生物型。的检测限之间的测定（16-1600 fg）表明，一些测定不应该被应用于临床样品，但可能有助于确定菌落material.Conclusions：总之，大多数的测定显示低检测限，并证明是高度选择性的布鲁氏菌属和物种的检测是最相关的人类。可以推荐使用靶向bcsp 31基因的检测来筛查布鲁氏菌。应连续应用种属特异性检测，通过第二个基因靶标确认初步诊断。
Background: The identification of Brucella isolates using conventional microbiological techniques is time-consuming and hazardous. We therefore assessed the performance of real-time PCR assays for the identification of members of the genus Brucella to the genus and species level.Methods: We evaluated an in-house developed assay and various previously published real-time PCR assays targeting bcsp31, per, IS711, alkB/IS711 and BMEI1162/IS711 using 248 Brucella strains representing the biotypes of all species and a large panel of clinically relevant, phylogenetically related and serologically cross-reacting bacteria.Results: No misidentification was observed. However, several biotypes of Brucella abortus and Brucella suis were not detected with some of the published assays. The limit of detection varied widely among the assays (16-1600 fg) demonstrating that some assays should not be applied to clinical samples but may help to identify colony material.Conclusions: In summary, most of the assays revealed low detection limits and proved to be highly selective for the detection of the genus Brucella and the species that are most relevant for humans. Assays targeting the bcsp31 gene can be recommended to screen for Brucella. Species-specific assays should be consecutively applied confirming the primary diagnosis by a second gene target.