Molecular serotyping of diarrheagenic Escherichia coli with a MeltArray assay reveals distinct correlation between serotype and pathotype.

IF 12.2 1区 医学 Q1 GASTROENTEROLOGY & HEPATOLOGY Gut Microbes Pub Date : 2024-09-18 DOI:10.1080/19490976.2024.2401944
Chen Du,Yiqun Liao,Congcong Ding,Jiayu Huang,Shujuan Zhou,Yiyan Xu,Zhaohui Yang,Xiaolu Shi,Yinghui Li,Min Jiang,Le Zuo,Minxu Li,Shengzhe Bian,Na Xiao,Liqiang Li,Ye Xu,Qinghua Hu,Qingge Li
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Abstract

Diarrheagenic Escherichia coli serotypes are associated with various clinical syndromes, yet the precise correlation between serotype and pathotype remains unclear. A major barrier to such studies is the reliance on antisera-based serotyping, which is culture-dependent, low-throughput, and cost-ineffective. We have established a highly multiplex PCR-based serotyping assay, termed the MeltArray E. coli serotyping (EST) assay, capable of identifying 163 O-antigen-encoding genes and 53 H-antigen-encoding genes of E. coli. The assay successfully identified serotypes directly from both simulated and real fecal samples, as demonstrated through spike-in validation experiments and a retrospective study. In a multi-province study involving 637 E. coli strains, it revealed that the five major diarrheagenic pathotypes have distinct serotype compositions. Notably, it differentiated 257 Shigella isolates into four major Shigella species, distinguishing them from enteroinvasive E. coli based on their distinct serotype profiles. The assay's universality was further corroborated by in silico analysis of whole-genome sequences from the EnteroBase. We conclude that the MeltArray EST assay represents a paradigm-shifting tool for molecular serotyping of E. coli, with potential routine applications for comprehensive serotype analysis, disease diagnosis, and outbreak detection.
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利用熔融阵列检测法对致泻性大肠埃希菌进行分子血清分型,发现血清型与病理型之间存在明显的相关性。
腹泻性大肠埃希菌血清型与各种临床综合征有关,但血清型与病理型之间的确切相关性仍不清楚。此类研究的一个主要障碍是依赖抗血清进行血清分型,这种方法依赖培养、通量低、成本效益差。我们建立了一种基于 PCR 的高度多重血清分型检测方法,称为 MeltArray 大肠杆菌血清分型(EST)检测方法,能够鉴定大肠杆菌的 163 个 O 抗原编码基因和 53 个 H 抗原编码基因。通过尖峰验证实验和一项回顾性研究,该测定法成功地从模拟和真实粪便样本中直接鉴定出血清型。在一项涉及 637 株大肠杆菌的多省研究中,它揭示了五种主要腹泻致病型具有不同的血清型组成。值得注意的是,它将 257 株志贺氏菌分离物分为四大志贺氏菌种,并根据其不同的血清型特征将它们与肠道入侵性大肠杆菌区分开来。对 EnteroBase 中的全基因组序列进行的硅分析进一步证实了该检测方法的普遍性。我们的结论是,MeltArray EST 检测法是大肠杆菌分子血清型分型的一种范式转换工具,有望常规应用于全面的血清型分析、疾病诊断和疫情检测。
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来源期刊
Gut Microbes
Gut Microbes Medicine-Microbiology (medical)
CiteScore
18.20
自引率
3.30%
发文量
196
审稿时长
10 weeks
期刊介绍: The intestinal microbiota plays a crucial role in human physiology, influencing various aspects of health and disease such as nutrition, obesity, brain function, allergic responses, immunity, inflammatory bowel disease, irritable bowel syndrome, cancer development, cardiac disease, liver disease, and more. Gut Microbes serves as a platform for showcasing and discussing state-of-the-art research related to the microorganisms present in the intestine. The journal emphasizes mechanistic and cause-and-effect studies. Additionally, it has a counterpart, Gut Microbes Reports, which places a greater focus on emerging topics and comparative and incremental studies.
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