Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin gene-specific loop-mediated isothermal amplification method.

Abstract

Reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) is a unique gene amplification method that can be completed within 35 min at 62.5 degrees C. In the present study, RT-LAMP was used to develop a rapid and sensitive laboratory diagnostic system for the H5N1 highly pathogenic avian influenza (HPAI). The sensitivity of the system was 0.1-0.01 plaque-forming units per reaction for HPAI-H5N1 viruses belonging to the genetically and antigenically distinct clade 1, represented by A/Vietnam/JP1203/2004, and clade 2, represented by A/Indonesia/JP283/2006. This RT-LAMP sensitivity is 10-fold higher than the sensitivity of standard one-step RT-PCR. By using viral RNAs extracted from avian influenza viruses of H1-H15 hemagglutinin (HA) subtypes and human pathogenic respiratory viruses, it was confirmed that the RT-LAMP system amplifies specifically RNA of the H5 subtype virus. The system detected H5-HA genes in throat swabs collected from humans as well as from wild birds. These results suggest that the present RT-LAMP system is a useful diagnostic tool for surveillance of recent outbreaks of the HPAI-H5N1 virus.

• Authors: Masaki Imai, Ai Ninomiya, Harumi Minekawa, Tsugunori Notomi, Toru Ishizaki, Phan Van Tu, Nguyen Thi Kim Tien, Masato Tashiro, Takato Odagiri
• Journal: Journal of virological methods (J Virol Methods) Vol. 141 Issue 2 Pg. 173-80 (May 2007) ISSN: 0166-0934 [Print] Netherlands
• PMID: 17218021 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Viral
• DNA Primers
• Hemagglutinin Glycoproteins, Influenza Virus
• hemagglutinin, avian influenza A virus

Topics

• Adolescent
• Adult
• Aged
• Animals
• Antigens, Viral (genetics)
• Base Sequence
• Child
• Child, Preschool
• Crows
• DNA Primers (genetics)
• Genes, Viral
• Hemagglutinin Glycoproteins, Influenza Virus (genetics)
• Humans
• Influenza A Virus, H5N1 Subtype (genetics, isolation & purification)
• Influenza A virus (genetics, isolation & purification)
• Influenza in Birds (diagnosis, virology)
• Influenza, Human (diagnosis, virology)
• Middle Aged
• Molecular Sequence Data
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Sensitivity and Specificity
• Species Specificity