开发用于检测和区分 mpox 病毒支系的多重液滴数字 PCR 方法。

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-11-26 DOI:10.1016/j.jviromet.2024.115078

Xiaoyue Chu , Hailong Chen , Rui Wu , Linghao Zhang , Yong Zhang , Hua Xu , Chaofeng Ma

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引用次数: 0

摘要

背景：世界卫生组织已宣布目前爆发的痘病毒疫情为国际关注的突发公共卫生事件。然而，将麻风痘病毒（MPXV）感染症状与其他正痘病毒区分开来的方法并不典型，因此有必要进行实验室确证试验来帮助临床诊断。因此，必须快速准确地检测和区分 MPXV 的各个支系：目的：开发了一种多重液滴数字 PCR（ddPCR）方法来检测和区分 MPXV 的不同支系，随后通过对 17 份临床样本的分析评估了该方法的灵敏度和易用性：方法：通过比较多种正痘病毒的完整基因组，设计了用于多重 ddPCR 的引物和探针。在 Biorad QX200 平台上对引物和探针浓度、反应条件进行了初步优化。检测了 17 份 MPXV 临床样本，并通过 Sanger 测序进行了验证：结果：所建立的 ddPCR 方法可以检测和区分 MPXV，其结果与 Sanger 测序的结果一致：结论：多重 ddPCR 可用于快速、准确地检测和区分 MPXV 的不同支系。

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Development of a multiplex droplet digital PCR method for detection and differentiation of mpox virus clades

Background

The current outbreak of mpox has been declared a public health emergency of international concern by the World Health Organization. However, distinguishing symptoms of mpox virus (MPXV) infection from other orthopoxviruses is atypical, necessitating laboratory confirmatory tests to aid in clinical diagnosis. Therefore, rapid and accurate detection and differentiation of various clades of MPXV are imperative.

Objective

A multiplex droplet digital PCR (ddPCR) method was developed to detect and differentiate various clades of MPXV with subsequent evaluation of its sensitivity and accessibility through the analysis of 17 clinical samples.

Methods

Primers and probes for multiple ddPCR were designed by comparing multiple complete genomes of orthopoxviruses. Primer and probe concentrations, reaction conditions were tentatively optimized on the Biorad QX200 platform. Seventeen clinical samples of MPXV were detected and verified by Sanger sequencing.

Results

The established ddPCR method could detect and differentiate MPXV, and the results were consistent with those of Sanger sequencing.

Conclusion

Multiplex ddPCR could be used to detect and distinguish different clades of MPXV rapidly and accurately.

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.