A one-step reverse-transcription recombinase aided PCR assay for the rapid and sensitive detection of human enteroviruses

• Main Authors: Xiuli Sun (Author), Huanhuan Lu (Author), Yanqing Tie (Author), Mengchuan Zhao (Author), Ruiqing Zhang (Author), Zhenlu Sun (Author), Guohao Fan (Author), Fengyu Li (Author), Fengyu Tian (Author), Yaxin Hu (Author), Mengyi Zhang (Author), Xinxin Shen (Author), Xuejun Ma (Author), Zhishan Feng (Author)
• Format: Book
• Published: Elsevier, 2023-04-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

Human enteroviruses (HEVs) include many different types that cause a wide range of diseases, and an effective method of genus-level identification has therefore significant clinical implications. However, quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), the gold-standard method, still has shortfalls in diagnostic sensitivity and timeliness. Here we established a one-step real-time reverse-transcription recombinase-aided PCR assay (RT-RAP) to detect HEV fragment within an hour. The RT-RAP assay showed a detection limit of 5 copies/μL using recombinant plasmids and was extensively verified using 15 HEV strains. Among 15 types of HEV (species A-C), the sensitivity of RT-RAP was approximately 2-8 folds lower than that of the qRT-PCR in 9 types, and no-cross reaction with other viruses was observed. RT-RAP was further applied to analyze CSF and fecal specimens; the clinical performance demonstrated that the RT-RAP and the commercial qRT-PCR kit provided consistent results. These results indicated that RT-RAP assay may be a promising approach for rapid and sensitive detection of HEV.

Item Description

2590-0536
10.1016/j.bsheal.2023.03.002