Rapid detection of human mastadenovirus species B by recombinase polymerase amplification assay

Tao Wu,Kangchen Zhao,Fengcai Zhu,Haizhen Wu,Minghao Zhou,Xingchen Zhang,Lunbiao Cui,Chaoyou Hu,Yiyue Ge,Xiaojuan Zhu

doi:10.1186/s12866-018-1365-7

Abstract

BackgroundAs an important component of the causative agent of respiratory tract infections, enteric and eye infections, Human mastadenoviruses (HAdVs) species B spread easily in the crowd. In this study, we developed a recombinase polymerase amplification (RPA) assay for rapidly detecting HAdVs species B which was comprised of two different formats (real-time and lateral-flow device).ResultsThis assay was confirmed to be able to detect 5 different HAdVs species B subtypes (HAdV-B3, HAdV-B7, HAdV-B11, HAdV-B14 and HAdV-B55) without cross-reactions with other subtypes and other respiratory tract pathogens. This RPA assay has not only highly sensitivity with low detection limit of 50 copies per reaction but also short reaction time (< 15 min per detection). Furthermore, the real-time RPA assay has excellent correlation with real-time PCR assay for detection of HAdVs species B presented in clinical samples.ConclusionsThus, the RPA assay developed in this study provides an effective and portable approach for the rapid detection of HAdVs species B.

Highlights

As an important component of the causative agent of respiratory tract infections, enteric and eye infections, Human mastadenoviruses (HAdVs) species B spread in the crowd
There were no significant differences in the lateral flow device (LFD)-recombinase polymerase amplification (RPA) and real-time RPA results for amplification at different temperature
The results revealed HAdVs species B could be detected by both LFD RPA and real-time RPA assays while other species could not be detected (Fig. 2)

Summary

Introduction

As an important component of the causative agent of respiratory tract infections, enteric and eye infections, Human mastadenoviruses (HAdVs) species B spread in the crowd. We developed a recombinase polymerase amplification (RPA) assay for rapidly detecting HAdVs species B which was comprised of two different formats (real-time and lateral-flow device). Immunological assay, such (2019) 19:8 as direct immunofluorescence, was rapid to detect viruses This assay usually did not show enough sensitivity and needed special antiserum. With the progress in molecular detection techniques, PCR or real-time PCR assays have rendered rapid and sensitive tools for detection, typing, and monitoring of adenoviral infections [12,13,14,15]. We present a recently developed isothermal amplification method, named recombinase polymerase amplification (RPA), for detecting HAdVs species B. To resolve the need for resourcelimited settings, portable real-time fluorescence scanner device and lateral flow device (LFD) were applied to detect the RPA products in present study

Methods

Results

Discussion

Conclusion