Journal
ARTIFICIAL CELLS NANOMEDICINE AND BIOTECHNOLOGY
Volume 51, Issue 1, Pages 233-240Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/21691401.2023.2203198
Keywords
Hepatitis A virus; reverse transcription multienzyme isothermal rapid amplification; lateral flow strip; nucleic acid detection; rapid diagnosis
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A feasible HAV detection solution was established by combining RT-MIRA and LFD. The RT-MIRA-LFD method showed high accuracy and rapidness in detecting HAV infection. This method could provide a considerable advantage for diagnosing and controlling HAV infection, especially in regions with limited medical resources.
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 degrees C and naked-eye observation of the LFD strips in 10 min. The detection sensitivity of this method reached 1 copy/mu l. 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.
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