Development of a reverse transcription loop-mediated isothermal amplification assay for the rapid diagnosis of avian influenza A (H7N9) virus infection
•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...
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| Published in | Journal of virological methods Vol. 204; pp. 101 - 104 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
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Elsevier B.V
01.08.2014
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| Abstract | •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. |
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| AbstractList | 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 42.47 copies/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 30 min, the RT-LAMP assay developed in this study is a promising rapid diagnostic tool for avian influenza A (H7N9) virus infection.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 42.47 copies/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 30 min, the RT-LAMP assay developed in this study is a promising rapid diagnostic tool for avian influenza A (H7N9) virus infection. 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 42.47 copies/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 30 min, the RT-LAMP assay developed in this study is a promising rapid diagnostic tool for avian influenza A (H7N9) virus infection. •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. 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. |
| Author | Takahashi, Hitoshi Nakauchi, Mina Kageyama, Tsutomu Takayama, Ikuyo Tashiro, Masato |
| Author_xml | – sequence: 1 givenname: Mina surname: Nakauchi fullname: Nakauchi, Mina – sequence: 2 givenname: Ikuyo surname: Takayama fullname: Takayama, Ikuyo – sequence: 3 givenname: Hitoshi surname: Takahashi fullname: Takahashi, Hitoshi – sequence: 4 givenname: Masato surname: Tashiro fullname: Tashiro, Masato – sequence: 5 givenname: Tsutomu surname: Kageyama fullname: Kageyama, Tsutomu email: tkage@nih.go.jp |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24747008$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | 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 |
| Title | Development of a reverse transcription loop-mediated isothermal amplification assay for the rapid diagnosis of avian influenza A (H7N9) virus infection |
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