用于检测和区分猴痘病毒分支的多重液滴数字PCR方法的开发
Development of a multiplex droplet digital PCR method for detection and differentiation of mpox virus clades.
作者信息
Chu Xiaoyue, Chen Hailong, Wu Rui, Zhang Linghao, Zhang Yong, Xu Hua, Ma Chaofeng
机构信息
Shaanxi Blood Center, Xi'an, Shaanxi 710061, PR China.
Xi'an Center for Disease Control and Prevention, Xi'an, Shaanxi 710054, PR China.
出版信息
J Virol Methods. 2025 Feb;332:115078. doi: 10.1016/j.jviromet.2024.115078. Epub 2024 Nov 26.
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.
背景
世界卫生组织已宣布当前的猴痘疫情为国际关注的突发公共卫生事件。然而，猴痘病毒（MPXV）感染的症状与其他正痘病毒的症状难以区分，这就需要实验室确诊检测来辅助临床诊断。因此，快速、准确地检测和区分MPXV的各个分支至关重要。
目的
开发一种多重液滴数字PCR（ddPCR）方法，用于检测和区分MPXV的各个分支，并通过对17份临床样本的分析评估其敏感性和可及性。
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平台上初步优化了引物和探针浓度、反应条件。对17份MPXV临床样本进行检测，并通过桑格测序进行验证。
结果
所建立的ddPCR方法能够检测和区分MPXV，结果与桑格测序结果一致。
结论
多重ddPCR可用于快速、准确地检测和区分MPXV的不同分支。
Zotero 插件