Development of a reverse transcription loop-mediated isothermal amplification assay for the rapid diagnosis of avian influenza A (H7N9) virus infection

• Published in: Journal of virological methods Vol. 204; pp. 101 - 104
• Main Authors: Nakauchi, Mina, Takayama, Ikuyo, Takahashi, Hitoshi, Tashiro, Masato, Kageyama, Tsutomu
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.08.2014
• Subjects: Animals; avian influenza; Avian influenza virus; Birds; cross reaction; diagnostic techniques; H7N9; Humans; Influenza A virus; Influenza A Virus, H7N9 Subtype - genetics; Influenza A Virus, H7N9 Subtype - isolation & purification; Influenza B virus; Influenza in Birds - diagnosis; Influenza in Birds - virology; Influenza, Human - diagnosis; Influenza, Human - virology; Nucleic Acid Amplification Techniques - methods; Rapid diagnosis; Reverse Transcription; reverse transcription loop-mediated isothermal amplification; RT-LAMP; Sensitivity and Specificity; Time; transcription (genetics); viruses; Zoonoses - diagnosis; Zoonoses - virology; Rapid diagnosis; RT-LAMP; Avian influenza virus; H7N9
• ISSN: 0166-0934; 1879-0984; 1879-0984
• DOI: 10.1016/j.jviromet.2014.03.028

•The reverse transcription-loop-mediated isothermal amplification assay was developed to detect avian influenza A (H7N9) virus epidemic in China.•The developed assays were evaluated using in vitro transcribed control RNA, clinical specimens, and clinical isolates.•The assays were shown to have high sensitivity and high specificity. A genetic diagnosis system for detecting avian influenza A (H7N9) virus infection using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) technology was developed. The RT-LAMP assay showed no cross-reactivity with seasonal influenza A (H3N2 and H1N1pdm09) or influenza B viruses circulating in humans or with avian influenza A (H5N1) viruses. The sensitivity of the RT-LAMP assay was 42.47copies/reaction. Considering the high specificity and sensitivity of the assay for detecting the avian influenza A (H7N9) virus and that the reaction was completed within 30min, the RT-LAMP assay developed in this study is a promising rapid diagnostic tool for avian influenza A (H7N9) virus infection.