Development and evaluation of a multiplex digital PCR method for sensitive and accurate detection of respiratory pathogens in children

Xiangpeng Chen; Fang Wang; Yiliang Fu; Luci Huang; Fei Li; Hongwei Zhao; Xiaolei Guan; Qiuping Li; Qi Li; Yilu Wang; Yong Guo; Zhengde Xie

doi:10.1016/j.virol.2023.109948

Development and evaluation of a multiplex digital PCR method for sensitive and accurate detection of respiratory pathogens in children

Virology. 2024 Feb:590:109948. doi: 10.1016/j.virol.2023.109948. Epub 2023 Nov 29.

Authors

Xiangpeng Chen¹, Fang Wang², Yiliang Fu¹, Luci Huang¹, Fei Li¹, Hongwei Zhao¹, Xiaolei Guan¹, Qiuping Li¹, Qi Li¹, Yilu Wang², Yong Guo³, Zhengde Xie⁴

Affiliations

¹ Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Research Unit of Critical Infection in Children, Chinese Academy of Medical Sciences, 2019RU016, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing, China.
² Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing, China.
³ Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing, China. Electronic address: [email protected].
⁴ Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Research Unit of Critical Infection in Children, Chinese Academy of Medical Sciences, 2019RU016, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing, China. Electronic address: [email protected].

PMID: 38064870
DOI: 10.1016/j.virol.2023.109948

Abstract

The emergence of multiplex digital polymerase chain reaction (dPCR) and other detection technologies for respiratory pathogens in recent years has facilitated greater understanding of respiratory virus epidemics. In this study, a multiplex dPCR method was developed and evaluated as a means of detecting five respiratory pathogens in children with acute lower respiratory tract infection (ALRTI). With 139 nasopharyngeal swabs collected from children with ALRTI, pathogens were detected using dPCR and quantitative real-time PCR (qPCR) methods. Of those specimens, dPCR detected 86 positive cases, while qPCR identified 84. Moreover, dPCR exhibited higher sensitivity than qPCR, and displayed no cross-reactivity with common respiratory pathogens. These findings suggest that dPCR-based method could become one of the most promising options for acute respiratory pathogen detection.

Keywords: Acute lower respiratory tract inflection; Multiplex digital PCR; Quantitative real-time PCR; Respiratory pathogen.

MeSH terms

Child
Humans
Multiplex Polymerase Chain Reaction / methods
Real-Time Polymerase Chain Reaction / methods
Respiratory Tract Infections* / diagnosis
Viruses* / genetics