Rapid detection and typing of influenza A and B by loop-mediated isothermal amplification: comparison with immunochromatography and virus isolation

Journal of Virological Methods
Masahiro ItoYoshinobu Okuno

Abstract

A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was established for the detection of influenza A virus H1 and H3 subtype strains and influenza B virus strains specifically. The total procedure from RNA extraction to virus typing was completed within 3h. In terms of specificity, the representative AH1, AH3 and B strains were detected only by strain-specific primers respectively. No cross-detection was observed. In terms of sensitivity, virus was detected at a minimum concentration of 10 ffu/ml. Eighty-three nasopharyngeal aspirates obtained from children diagnosed clinically with influenza were tested by the RT-LAMP assay, along with commercially available immunochromatography rapid diagnostic tests and by virus isolation. Virus was isolated from 78 samples (94%) and the subtype was determined by the hemagglutination inhibition test. Although it took at least 3 days, the detection sensitivity was the best of the three methods. With two rapid assays, the detection sensitivity of the RT-LAMP assay (85.5%) was higher than that of immunochromatography tests (75.9%). In addition, the RT-LAMP assay can be used to differentiate emerging influenza virus subtypes by selecting appropriate primer sets.

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Citations

Jun 5, 2012·Frontiers of Medicine·Albert Cheung-Hoi YuLok-Ting Lau
Apr 10, 2012·Molecular Biotechnology·Francesca SidotiRossana Cavallo
May 21, 2013·Molecular Diagnosis & Therapy·Anthony P Malanoski, Baochuan Lin
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Oct 1, 2008·Expert Opinion on Medical Diagnostics·Masayuki SaijoIchiro Kurane
Nov 7, 2009·Clinics in Laboratory Medicine·Wenjuan Wu, Yi-Wei Tang
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Jul 30, 2014·Journal of Virological Methods·Yong Kwan KimDong-Jun An
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Jan 14, 2022·Health Security·Corey R HartRajesh R Naik

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