A multiplex real-time RT-PCR assay for the detection of H1, H2 and H3 subtype avian influenza viruses

Avian influenza viruses (AIVs) are influenza A viruses, of which subtypes H1, H2 and H3 are highly transmissible in poultry and have the risk of transmission to human as well. It is important to establish an accurate, sensitive and convenient means of virus detection. In this study, we developed a m...

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Published inVirus genes Vol. 59; no. 2; pp. 333 - 337
Main Authors Yan, Sijing, Yang, Fan, Yao, Hangping, Dong, Dalu, Wu, Danna, Wu, Nanping, Ye, Chunsheng, Wu, Haibo
Format Journal Article
LanguageEnglish
Published New York Springer US 01.04.2023
Springer Nature B.V
Subjects
Animals
Avian flu
Biomedical and Life Sciences
Biomedicine
Brief Report
Genes
Hemagglutinin Glycoproteins, Influenza Virus - genetics
Hemagglutinins
Humans
Infectious diseases
Influenza A
Influenza A virus - genetics
Influenza in Birds - diagnosis
Laboratories
Medical diagnosis
Medical Microbiology
Medical research
Orthomyxoviridae
Pandemics
Plant Sciences
Plasmids
Polymerase chain reaction
Poultry
Reproducibility
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Sensitivity and Specificity
Swine flu
Virology
Viruses
Multiplex PCR
Virus detection
Fluorescent quantitative PCR
Avian influenza virus (AIV)
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Summary:Avian influenza viruses (AIVs) are influenza A viruses, of which subtypes H1, H2 and H3 are highly transmissible in poultry and have the risk of transmission to human as well. It is important to establish an accurate, sensitive and convenient means of virus detection. In this study, we developed a multiplex real-time RT-PCR assay based on conserved sequences of the virus hemagglutinin and matrix, and designed primers and probes for the simultaneous and rapid detection of AIV subtypes H1, H2 and H3. We used different subtypes of AIVs and other avian respiratory viruses for evaluation of the specificity of this method. The results showed good sensitivity, specificity and reproducibility. The detection limit was 10–100 copies per reaction. The method also achieved good concordance with the virus isolation method when compared to 81 poultry samples evaluated. It provides a new method for detecting mixed infections of AIVs.
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ISSN:0920-8569
1572-994X
DOI:10.1007/s11262-022-01963-z