Visual detection of human infection with influenza A (H7N9) virus by subtype-specific reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye.

Abstract

A rapid and sensitive H7 and N9 subtype-specific reverse transcription loop-mediated isothermal amplification assay was developed respectively for visual detection of human-infected influenza A (H7N9) virus. The reaction was performed in one step in a single tube at 63°C for 60 min with the addition of hydroxynaphthol blue dye before amplification. The detection limits of both subtype-specific assays were comparable to those of validated H7 and N9 real-time PCR assays respectively and no cross-detection was observed with influenza A pandemic H1N1, H3N2, H5N1, H9N2 or influenza B virus. The assays were evaluated further with H7N9 virus-infected clinical specimens.

• Authors: K Nie, X Zhao, X Ding, X D Li, S M Zou, J F Guo, D Y Wang, R B Gao, X Y Li, W J Huang, Y L Shu, X J Ma
• Journal: Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases (Clin Microbiol Infect) Vol. 19 Issue 8 Pg. E372-5 (Aug 2013) ISSN: 1469-0691 [Electronic] England
• PMID: 23718218 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

Chemical References

• Naphthalenesulfonates
• trisodium 3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate

Topics

• Animals
• Humans
• Influenza A Virus, H7N9 Subtype (isolation & purification)
• Influenza, Human (diagnosis)
• Molecular Diagnostic Techniques (methods)
• Naphthalenesulfonates (metabolism)
• Nucleic Acid Amplification Techniques (methods)
• Sensitivity and Specificity
• Staining and Labeling (methods)
• Temperature
• Virology (methods)