Multiplex, bead-based suspension array for molecular determination of common Salmonella serogroups
Multiplex, bead-based suspension array for molecular determination of common Salmonella serogroups
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DOI：
10.1128/jcm.00025-07
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发表时间：
2007-10-01
影响因子：
9.4
通讯作者：
Fields, Patricia
中科院分区：
文献类型：
--
作者：
Fitzgerald, Collette;Collins, Marcus;Fields, Patricia
我们报告了沙门氏菌O群特异性生物检测试剂盒的开发和评价，该试剂盒用于检测美国最常见的六种血清群（B、C-1、C-2、D、E和013）以及甲型副伤寒血清型。该测定基于直接参与O-抗原生物合成的rfb基因靶标;它可以在PCR扩增后45分钟完成。与传统的血清分型相比，该检测试剂正确且特异性地鉴定了384株（94.3%）分离株。17株（4.4%）产生的结果与已知的血清型的分子基础一致，但与传统的血清分型结果不同，5株（1.3%）产生假阴性结果。在大多数情况下，粗糙分离株血清型的分子测定与常见血清型一致，表明这种方法有可能为不表达O抗原的分离株提供O型信息。我们还报告了肠道沙门氏菌Poona血清型（血清群O 13）O-抗原编码rfb基因簇的序列。与其他，以前的特点rfb区域相比，O 13 rfb基因簇是最密切相关的大肠杆菌O 127和086。本文所述的O-群生物检测试剂盒为快速检测常见沙门氏菌血清群提供了一种易于使用的高通量系统。
We report the development and evaluation of a Salmonella O-group-specific Bio-Plex assay to detect the six most common serogroups in the United States (B, C-1, C-2, D, E, and 013) plus serotype Paratyphi A. The assay is based on rfb gene targets directly involved in O-antigen biosynthesis; it can be completed 45 min post-PCR amplification. The assay correctly and specifically identified 362 of 384 (94.3%) isolates tested in comparison to traditional, serotyping. Seventeen isolates (4.4%) produced results consistent with what is known about the molecular basis for serotypes but different from the results of traditional serotyping, and five isolates (1.3%) generated false-negative results. Molecular determination of the serogroup for rough isolates was consistent with a common serotype in most instances, indicating that this approach has the potential to provide O-group information for isolates that do not express an O antigen. We also report the sequence of the O-antigenen-coding rfb gene cluster from Salmonella enterica serotype Poona (serogroup O13). Compared with other, previously characterized rfb regions, the O13 rfb gene cluster was most closely related to Escherichia coli O127 and 086. The O-group Bio-Plex assay described here provides an easy-to-use, high-throughput system for rapid detection of common Salmonella serogroups.