Single-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) for influenza A virus subtype H5N1 detection
Single-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) for influenza A virus subtype H5N1 detection
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DOI：
10.1089/vim.2004.17.588
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发表时间：
2004-12-01
期刊：
影响因子：
2.2
通讯作者：
Poovorawan, Y
中科院分区：
文献类型：
--
作者：
Payungporn, S;Phakdeewirot, P;Poovorawan, Y
甲型流感病毒 H5N1 亚型在家禽中引起一种快速致命的全身性疾病，并直接从家禽传播给人类。本研究的目的是开发一种快速、节省成本且有效的甲型流感病毒 H5N1 亚型检测方法。选定的引物组用于单步 RT-PCR，以多重形式同时检测 276、189 和 131 bp 片段，对应于 M、H5 和 N1 的特异性序列。扩增的DNA片段通过琼脂糖凝胶电泳清晰分离。该测定的灵敏度约为10(3)copies/μL。此外，该方法不仅可用于检测禽类，还可用于检测人类甲型流感病毒H5N1亚型。总之，这种特殊方法的亮点在于其快速性和成本效益，因此对于甲型流感病毒H5N1亚型爆发时的大规模筛查具有可行性和吸引力。
Influenza A virus subtype H5N1 causes a rapidly fatal systemic disease in domestic poultry and spreads directly from poultry to humans. The aim of this study was to develop a rapid, cost-saving and effective method for influenza A virus subtype H5N1 detection. The selected primer set was used in single-step RT-PCR for simultaneous detection in multiplex format of the 276-, 189-, and 131-bp fragments, corresponding to sequences specific for M, H5 and N1. The amplified DNA fragments were clearly separated by agarose gel electrophoresis. The sensitivity of this assay was about 10(3)copies/muL. Moreover, this method can be applied to detect not only avian but also human influenza A virus subtype H5N1. In conclusion, the highlights of this particular method are its rapidity and cost-effectiveness, thus rendering it feasible and attractive for large-scale screening at times of influenza A virus subtype H5N1 outbreak.