Broad Detection of Diverse H5 and H7 Hemagglutinin Genes of Avian Influenza Viruses by Real-Time Reverse Transcription-PCR Using Primer and Probe Sets Containing Mixed Bases

• Published in: Journal of Clinical Microbiology Vol. 48; no. 11; pp. 4275 - 4278
• Main Authors: Tsukamoto, Kenji, Noguchi, Daigo, Suzuki, Koutaro, Shishido, Makiko, Ashizawa, Takayoshi, Kim, Min-Chul, Lee, Youn-Jeong, Tada, Tatsuya
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-11-2010; American Society for Microbiology (ASM)
• Subjects: Animals; Biological and medical sciences; Birds; DNA Primers - genetics; Fundamental and applied biological sciences. Psychology; Hemagglutinin Glycoproteins, Influenza Virus - genetics; Hemagglutinin Glycoproteins, Influenza Virus - isolation & purification; Influenza in Birds - diagnosis; Influenza in Birds - virology; Microbiology; Miscellaneous; Molecular Sequence Data; Oligonucleotide Probes - genetics; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Viral - genetics; Sensitivity and Specificity; Sequence Analysis, DNA; Virology; Virology - methods; Virus; Influenzavirus A; Gene; Avian influenzavirus; Hemagglutinin; Orthomyxoviridae; Zoopathogen; Detection; Real time; Reverse transcription polymerase chain reaction
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.01264-10

Real-time reverse transcription-PCR (RT-PCR) was developed for broad detection of diverse H5 and H7 genes in Eurasian and American lineages of avian influenza viruses by using primer and probe sets containing mixed bases. Optimal use of the mixed bases enabled us to minimize sequence mismatches and to broaden the gene detection spectrum without decreasing sensitivity.