Visual detection of H1 subtype and identification of N1, N2 subtype of avian influenza virus by reverse transcription loop-mediated isothermal amplification assay

In order to visually detect H1, N1 and N2 subtype of avian influenza virus (AIV), three reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed. According to the sequences of AIV gene available in GenBank, three degenerate primer sets specific to HA gene of H1 su...

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Bibliographic Details
Published inBingdu xuebao Vol. 29; no. 2; p. 154
Main Authors Peng, Yi, Xie, Zhi-Xun, Guo, Jie, Zhou, Chen-Yu, Liu, Jia-Bo, Pang, Yao-Shan, Deng, Xian-Wen, Xie, Zhi-Qin, Xie, Li-Ji, Fan, Qing, Luo, Si-Si
Format Journal Article
LanguageChinese
Published China 01.03.2013
Subjects
Animals
Chickens
DNA Primers - genetics
Ducks
Influenza A virus - classification
Influenza A virus - genetics
Influenza A virus - isolation & purification
Influenza A Virus, H1N1 Subtype - classification
Influenza A Virus, H1N1 Subtype - genetics
Influenza A Virus, H1N1 Subtype - isolation & purification
Influenza A Virus, H1N2 Subtype - classification
Influenza A Virus, H1N2 Subtype - genetics
Influenza A Virus, H1N2 Subtype - isolation & purification
Influenza in Birds - diagnosis
Influenza in Birds - virology
Nucleic Acid Amplification Techniques - methods
Poultry Diseases - diagnosis
Poultry Diseases - virology
Reverse Transcription
Turkeys
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Summary:In order to visually detect H1, N1 and N2 subtype of avian influenza virus (AIV), three reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed. According to the sequences of AIV gene available in GenBank, three degenerate primer sets specific to HA gene of H1 subtype AIV, NA gene of N1 and N2 subtype AIV were designed, and the reaction conditions were optimized. The results showed that all the assays had no cross-reaction with other subtype AIV and other avian respiratory pathogens, and the detection limit was higher than that of conventional RT-PCR. These assays were performed in water bath within 50 minutes. Without opening tube, the amplification result could be directly determined by inspecting the color change of reaction system as long as these assays were fin-ished. Fourteen specimens of H1N1 subtype and eight specimens of H1N2 subtype of AIV were identified from the 120 clinical samples by RT-LAMP assays developed, which was consistent with that of virus isolation
ISSN:1000-8721