Simple and feasible detection of hepatitis a virus using reverse transcription multienzyme isothermal rapid amplification and lateral flow dipsticks without standard PCR laboratory

• Main Authors: Li, X.-N (Author), Pan, Y.-Q (Author), Sun, M.-L (Author), Yao, J. (Author), Zhong, Y. (Author)
• Format: Article
• Language: English
• Published: Taylor and Francis Ltd. 2023
• Subjects: Diagnosis; genetics; Hepatitis A virus; human; Humans; Isotherms; Lateral Flow; Lateral flow dipsticks; lateral flow strip; Lateral flow strip; nucleic acid detection; Nucleic acid detection; polymerase chain reaction; Polymerase chain reaction; Polymerase Chain Reaction; Rapid diagnose; rapid diagnosis; reverse transcription; Reverse transcription; Reverse Transcription; reverse transcription multienzyme isothermal rapid amplification; Reverse transcription multienzyme isothermal rapid amplification; RNA; sensitivity and specificity; Sensitivity and Specificity; Simple++; Virus infection; Viruses
• Online Access: View Fulltext in Publisher

 Hepatitis A virus (HAV) is mainly transmitted via contaminated food and water. HAV infection is a major global public health problem. Thus, developing a simple, rapid detection method is crucial for containing HAV epidemics, particularly in developing regions with limited laboratory resources. This study established a feasible HAV detection solution by combining reverse transcription multienzyme isothermal rapid amplification (RT-MIRA) and lateral flow dipstick (LFD) strips. Primers targeting the conserved 5’UTR sequence of HAV were used in the RT-MIRA-LFD assay. RNA extraction was enhanced by obtaining RNA directly from the centrifuged supernatant. Our study found that MIRA amplification could be finished in 12 min at 37 °C and naked-eye observation of the LFD strips in 10 min. The detection sensitivity of this method reached 1 copy/μl. RT-MIRA-LFD was compared to conventional RT-PCR using 35 human blood samples. The accuracy of the RT-MIRA-LFD method was 100%. The convenience, sensitivity, and rapidness of this detection method could provide a considerable advantage for diagnosing and controlling HAV infection, especially in regions with limited medical resources. © 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.