Rapid Serotyping of Salmonella Isolates Based on Single Nucleotide Polymorphism-Like Sequence Profiles of a Salmonella-Specific Gene
Rapid Serotyping of Salmonella Isolates Based on Single Nucleotide Polymorphism-Like Sequence Profiles of a Salmonella-Specific Gene
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基于沙门氏菌特异性基因的单核苷酸多态性样序列图谱对沙门氏菌分离株进行快速血清分型
DOI：
10.1089/fpd.2020.2823
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发表时间：
2021
影响因子：
2.8
通讯作者：
Hisao Kurazono
中科院分区：
文献类型：
--
作者：
Eiki Yamasaki;Shigeru Matsuzawa;Kaoru Takeuchi;Yo Morimoto;Tetsuya Ikeda;Kayo Okumura;Hisao Kurazono
虽然血清分型是沙门氏菌最重要的分类鉴定方法，但常规的血清分型需要一套完整的抗血清，费时费力。最近，已经开发了聚合酶链反应（PCR）的快速血清分型程序。在这项研究中，我们建立了一种新的基于PCR的快速血清分型方法，采用了一个独特的目标基因。对沙门氏菌特异性基因（Salmonellaenterotoxin）的序列比对研究表明，该基因序列与沙门氏菌的血清型相关。在stn基因的750 bp序列中，有55个核苷酸呈现单核苷酸多态性（single nucleotide polymorphism，SNP）样多态性，且SNP样多态性与血清型的相关性表明，stn基因的SNP样序列可作为血清型的一个指标。为了建立一种基于SNP-like多态性的快速血清学分型方法，我们选择了stn基因上与SNP相关的12个SNP-like位点，建立了一种基于高分辨熔解（HRM）和PCR的方法，该方法在1.5 h内鉴定出SNP位点的核苷酸。这种新建立的快速血清分型程序（stn-HRM）可以识别9种血清型，包括经常分离的肠道菌血清型。根据世界卫生组织/全球食源性感染网络（WHO/GFN）国家数据库2001年至2010年的报告，这9种血清型涵盖了沙门氏菌病例的64.3%。在这项研究中，我们采用了一个独特的靶基因，p53，这是完全独立的基因，在以前报道的快速血清分型程序的目标。因此，我们新开发的stn-HRM程序所获得的结果是独立的其他程序所获得的结果。此外，stn-HRM可以确保细菌物种asstnis a沙门氏菌特异性基因的准确鉴定。新构建的stn-HRM与先前报道的方法相结合有望进一步提高沙门氏菌分离株血清分型的可信度。
Although serotyping is the most important method of identification of taxonomy inSalmonella, conventional serotype determination with a complete set of antisera is time consuming and laborious. Recently, rapid serotyping procedures with polymerase chain reaction (PCR) have been developed. In this study, we established a novel PCR-based rapid serotyping method that employs a unique target gene. Alignment study ofSalmonella-specific gene (Salmonellaenterotoxin [stn]) revealed a correlation between thestngene sequence and the serotype of the organism. In 750 bp ofstngene, 55 nucleotides indicated single nucleotide polymorphism (SNP)-like polymorphism, and the correlation between the SNP-like polymorphism and the serotype of the organism suggests that SNP-like sequences instngene can serve as an index for serotyping. To develop a rapid serotyping method based on the SNP-like polymorphism, we selected serotype-associated 12 SNP-like sites in thestngene and established a method based on high-resolution melting (HRM) and PCR, which identifies nucleotides at SNP-like sites within 1.5 h. This newly established rapid serotyping procedure (stn-HRM) could identify nine serotypes, including the frequently isolated serovar Enteritidis. These nine serotypes cover 64.3% of cases ofSalmonella, as reported by the World Health Organization/Global Foodborne Infection Network (WHO/GFN) Country Databank from 2001 to 2010. In this study, we employed a unique target gene,stn, which is completely independent of the genes that were targeted in previously reported rapid serotyping procedures. Therefore, the results obtained by our newly developedstn-HRM procedure are independent of the results obtained by other procedures. Besides,stn-HRM can ensure accurate identification of the bacterial species asstnis aSalmonella-specific gene. It is expected that the combination of newly constructedstn-HRM and previously reported procedures could further improve the credibility ofSalmonellaisolate serotyping.