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多重RT-RPA技术检测常见儿童腹泻病原体的方法研究

韩明月 陈坚 沈丽君 方丽娟 余道军

中国现代医生2026,Vol.64Issue(12):6-10,5.
中国现代医生2026,Vol.64Issue(12):6-10,5.DOI:10.3969/j.issn.1673-9701.2026.12.002

多重RT-RPA技术检测常见儿童腹泻病原体的方法研究

Research on the method of multiplex RT-RPA technology for detecting common pathogens causing diarrhea in children

韩明月 1陈坚 2沈丽君 1方丽娟 1余道军3

作者信息

  • 1. 杭州市第九医院检验科,浙江 杭州 311225
  • 2. 杭州市余杭区第一人民医院检验科,浙江 杭州 311113
  • 3. 西湖大学医学院附属杭州市第一人民医院医学检验科,浙江 杭州 310003
  • 折叠

摘要

Abstract

Objective To establish a reverse transcription recombinase polymerase amplification(RT-RPA)that can simultaneously detect multiple common pathogens causing diarrhea in children.Methods Conservative sequences of rotavirus,adenovirus,and norovirus genomes were selected to design recombinase polymerase amplification(RPA)primer and probe,and a multiplex RT-RPA detection system was established.The sensitivity,specificity,and repeatability of the established multiplex RT-RPA reaction system were validated,and small clinical sample validation was conducted on the established system.Results The multiplex RT-RPA established in this study has a detection sensitivity of 101copies/μl,102copies/μl and 102copies/μl for rotavirus,adenovirus,and norovirus,respectively,and there was no cross-reaction with other common viruses.Different concentrations of samples were tested 10 times,and the standard deviation of Ct values was small,with coefficients of variation less than 5%.There is no significant difference in the detection rate of common pediatric diarrhea pathogens in clinical samples between PCR method and the colloidal gold method.Conclusion The multiplex RT-RPA established in this study has good sensitivity,specificity,and reproducibility,and can provide new ideas for the detection of pathogens in childhood diarrhea.

关键词

轮状病毒/腺病毒/诺如病毒/多重RT-RPA

Key words

Rotavirus/Adenovirus/Norovirus/Multiplex RT-RPA

分类

医药卫生

引用本文复制引用

韩明月,陈坚,沈丽君,方丽娟,余道军..多重RT-RPA技术检测常见儿童腹泻病原体的方法研究[J].中国现代医生,2026,64(12):6-10,5.

基金项目

杭州市医药卫生科技项目(A20230151) (A20230151)

中国现代医生

1673-9701

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