Rapid and specific differentiation of Salmonella enterica serotypes typhi and Paratyphi by multicolor melting curve analysis.

IF 4.3 3区 医学 Q1 GASTROENTEROLOGY & HEPATOLOGY
Yixiang Jiang, Min Jiang, Rui Cai, Xiaolu Shi, Qinghua Hu, Biao Kan
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引用次数: 0

Abstract

Rapid and accurate identification of Salmonella enterica serotypes Typhi and Paratyphi (A, B and C), the causal agents of enteric fever, is critical for timely treatment, case management and evaluation of health policies in low and middle-income countries where the disease still remains a serious public health problem. The present study describes the development of a multiplex assay (EFMAtyping) for simultaneous identification of pathogens causing typhoid and paratyphoid fever in a single reaction by the MeltArray approach, which could be finished within 2.5 h. Seven specific genes were chosen for differentiation of typhoidal and nontyphoidal Salmonella. All gene targets were able to be detected by the EFMAtyping assay, with expected Tm values and without cross-reactivity to other relevant Salmonella serovars. The limit of detection (LOD) for all gene targets was 50 copies per reaction. The LOD reached 102-103 CFU/ml for each pathogen in simulated clinical samples. The largest standard deviation value for mean Tm was below 0.5 °C. This newly developed EFMAtyping assay was further evaluated by testing 551 clinical Salmonella isolates, corroborated in parallel by the traditional Salmonella identification workflow, and serotype prediction was enabled by whole-genome sequencing. Compared to the traditional method, our results exhibited 100% of specificity and greater than 96% of sensitivity with a kappa correlation ranging from 0.96 to 1.00. Thus, the EFMAtyping assay provides a rapid, high throughput, and promising tool for public health laboratories to monitor typhoid and paratyphoid fever.

通过多色熔解曲线分析法快速特异性区分伤寒沙门氏菌和副伤寒沙门氏菌血清型。
肠炎沙门氏菌血清型 Typhi 和 Paratyphi(A、B 和 C)是肠炎的致病菌,在肠炎仍是严重公共卫生问题的中低收入国家,快速准确地鉴定肠炎沙门氏菌血清型 Typhi 和 Paratyphi(A、B 和 C)对于及时治疗、病例管理和卫生政策评估至关重要。本研究介绍了一种多重检测方法(EFMAtyping)的开发情况,该方法采用熔融阵列(MeltArray)方法,可在 2.5 小时内完成一次反应,同时鉴定引起伤寒和副伤寒的病原体。所有基因靶标都能被 EFMA 分型检测法检测到,并具有预期的 Tm 值,且与其他相关沙门氏菌血清没有交叉反应。所有基因靶标的检测限(LOD)均为每个反应 50 个拷贝。在模拟临床样本中,每种病原体的检测限均达到 102-103 CFU/ml。平均 Tm 的最大标准偏差值低于 0.5 °C。新开发的 EFMA 分型测定通过检测 551 例临床沙门氏菌分离物进行了进一步评估,并与传统的沙门氏菌鉴定工作流程进行了平行印证,通过全基因组测序实现了血清型预测。与传统方法相比,我们的结果显示特异性为 100%,灵敏度超过 96%,卡帕相关性在 0.96 至 1.00 之间。因此,EFMA分型测定为公共卫生实验室监测伤寒和副伤寒提供了一种快速、高通量且前景广阔的工具。
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来源期刊
Gut Pathogens
Gut Pathogens GASTROENTEROLOGY & HEPATOLOGY-MICROBIOLOGY
CiteScore
7.70
自引率
2.40%
发文量
43
期刊介绍: Gut Pathogens is a fast publishing, inclusive and prominent international journal which recognizes the need for a publishing platform uniquely tailored to reflect the full breadth of research in the biology and medicine of pathogens, commensals and functional microbiota of the gut. The journal publishes basic, clinical and cutting-edge research on all aspects of the above mentioned organisms including probiotic bacteria and yeasts and their products. The scope also covers the related ecology, molecular genetics, physiology and epidemiology of these microbes. The journal actively invites timely reports on the novel aspects of genomics, metagenomics, microbiota profiling and systems biology. Gut Pathogens will also consider, at the discretion of the editors, descriptive studies identifying a new genome sequence of a gut microbe or a series of related microbes (such as those obtained from new hosts, niches, settings, outbreaks and epidemics) and those obtained from single or multiple hosts at one or different time points (chronological evolution).
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