Development of a DNA Microarray for Molecular Identification of All 46 Salmonella O Serogroups
Development of a DNA Microarray for Molecular Identification of All 46 Salmonella O Serogroups
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开发用于所有 46 种沙门氏菌 O 血清群分子鉴定的 DNA 微阵列
DOI：
10.1128/aem.00225-13
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发表时间：
2013-06-01
影响因子：
4.4
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Guo, Dan;Liu, Bin;Wang, Lei
摘要 沙门氏菌是许多国家食源性疾病的主要原因。沙门氏菌的血清型测定对于疾病评估、感染控制和流行病学监测具有重要意义。在这项研究中，开发了一种针对 O 抗原特异性基因的微阵列系统，用于同时检测和鉴定所有 46 个沙门氏菌 O 血清群。其中，40个血清群可以被可靠地鉴定出来，其余6个血清群分为三对（血清群O67和B、E1和E4、A和D1），需要使用PCR方法或常规血清分型方法进一步区分。当针对 293 种沙门氏菌菌株、186 种志贺氏菌菌株、代表性大肠杆菌菌株和 10 种其他细菌物种进行评估时，该微阵列显示出高度特异性。该检测正确鉴定了 288 种 (98%) 沙门氏菌菌株。检测灵敏度确定为每个样品 50 ng 基因组 DNA。通过在番茄背景下测试模拟样品，富集后每克接种可检测到 2 至 8 CFU。这种新开发的微阵列检测是第一个可用于全面检测和鉴定所有 46 个沙门氏菌 O 血清群的分子方案。与传统的血清分型方法相比，微阵列提供了一种可靠、高通量、灵敏的方法，可用于同时快速鉴定多个沙门氏菌 O 血清群。
ABSTRACT Salmonella is a major cause of food-borne disease in many countries. Serotype determination of Salmonella is important for disease assessment, infection control, and epidemiological surveillance. In this study, a microarray system that targets the O antigen-specific genes was developed for simultaneously detecting and identifying all 46 Salmonella O serogroups. Of these, 40 serogroups can be confidently identified, and the remaining 6, in three pairs (serogroups O67 and B, E1 and E4, and A and D1), need to be further distinguished from each other using PCR methods or conventional serotyping methods. The microarray was shown to be highly specific when evaluated against 293 Salmonella strains, 186 Shigella strains, representative Escherichia coli strains, and 10 strains of other bacterial species. The assay correctly identified 288 (98%) of the Salmonella strains. The detection sensitivity was determined to be 50 ng genomic DNA per sample. By testing simulated samples in a tomato background, 2 to 8 CFU per gram inoculated could be detected after enrichment. This newly developed microarray assay is the first molecular protocol that can be used for the comprehensive detection and identification of all 46 Salmonella O serogroups. Compared to the traditional serogrouping method, the microarray provides a reliable, high-throughput, and sensitive approach that can be used for rapid identification of multiple Salmonella O serogroups simultaneously.