Establishment and preliminary application of RPA-capillary electrophoresis for detection of human adenovirus 55
YUAN Ruodong
DONG Yangchao
WU Fuxing
DUAN Tian
XUE Pan
ZHANG Jian
YUAN Mingcheng
XUE Zhifeng
ZHANG Haijun
ZHANG Qianqian
LEI Yingfeng
GAO Xiaopeng
Abstract：Objective To establish a method for rapid detection of human adenovirus type 55(HAdV55)by recombinase polymerase amplification(RPA)combined with capillary electrophoresis and its preliminary application.Methods The HAdV55-Hexon plasmid was constructed as the amplification template,and RPA specific primers were designed for Hexon gene sequence of HAdV55 conserved region.The primers were screened and optimized,and the RPA amplification and combined capillary electrophoresis detection system was established to evaluate the specificity,sensitivity and reliability.Eighteen clinical samples were examined by RPA-capillary electrophoresis.Results A plasmid containing the full-length HAdV55-Hexon gene was constructed,and a pair of RPA primers with the highest amplification efficiency was obtained.The RPA-capillary electrophoresis detection system of HAdV55 had a minimum detection limit of 2.9 x 103 copies/mL,and the whole process could be completed in 30 min.The standard extracts of coronavirus(HCoV-229E,HCoV-OC43,and HCoV-HKU1),influenza A and B viruses,parainfluenza virus and respiratory syncytial virus were all negative when tested with this system.In addition,common human adenovirus types(HAdV3,HAdV4,HAdV7,HAdV16,and HAdV21)were also negative,which suggested that it had strong specificity for HAdV55.The established RPA-capillary electrophoresis assay could effectively detect HAdV55 in throat swabs of clinical patients.Conclusion The establishment of RPA combined with capillary electrophoresis for the detection of HAdV55 provides a good basis for its clinical diagnosis.
Keywords：human adenovirus type 55recombinase polymerase amplificationcapillary electrophoresis
Publication Date：2024-01-28
Online Publishing Date：2026-08-26(First online date of this platform, not the publication date of the document)
Pages：7( 66-72 )