[古巴甲型流感病毒（H1N1）分子检测方法的设计与实施]
[Design and implementation of a molecular method for influenza A virus (H1N1) in Cuba].
作者信息
Valdés Ramírez Odalys, Piñón Ramos Alexander, Acosta Herrera Belsy, Savón Valdés Clara, González Muñoz Grehete, Arencibia García Amely, Guilarte García Elías, González Báez Guelsys, Oropeza Fernández Suset, Hernández Espinosa Bárbara, Goyenechea Hernández Angel
机构信息
Departamento de Virología, Instituto de Medicina Tropical "Pedro Kouri" (IPK), La Habana, Cuba.
出版信息
Rev Cubana Med Trop. 2011 Jan-Apr;63(1):15-20.
INTRODUCTION
From March through April of 2009, Mexico notified outbreaks of respiratory illness, due to a new influenza virus of swine origin, which spread over rapidly via human-to-human transmission. The molecular methods currently in use were not suitable because the genome composition based on gene segments of swine, avian and human origin was quite different from the influenza A virus (H1N1) circulating at that time.
OBJECTIVE
Based on the published sequences, a set of specific primers for the HA gene was designed to evaluate a new RT-PCR assay.
METHODS
The RT-PCR assay processed 3 197 clinical samples from suspected cases of pandemic influenza A (H1N1) infection.
RESULTS
The novel optimized method obtained a 262 pb segment, without unspecific reactions. The new method proved to be useful in the diagnosis and subtyping of pandemic HINI influenza virus. The amplified product was verified by nucleotide sequencing, thus confirming the virus.
CONCLUSIONS
The introduction of this new assay for the laboratory surveillance of influenza virus strengthens the diagnostic capacity of the National Reference Laboratory.
引言
2009年3月至4月，墨西哥通报了由一种新型猪源流感病毒引起的呼吸道疾病疫情，该病毒通过人际传播迅速蔓延。目前使用的分子方法并不适用，因为基于猪、禽和人源基因片段的基因组组成与当时流行的甲型流感病毒（H1N1）有很大不同。
目的
基于已发表的序列，设计了一组针对HA基因的特异性引物，以评估一种新的逆转录聚合酶链反应（RT-PCR）检测方法。
方法
RT-PCR检测方法对3197份甲型H1N1流感疑似病例的临床样本进行了检测。
结果
新优化的方法获得了一个262个碱基对的片段，无非特异性反应。新方法在甲型H1N1流感大流行病毒的诊断和亚型鉴定中被证明是有用的。扩增产物经核苷酸测序验证，从而确认了病毒。
结论
这种用于流感病毒实验室监测的新检测方法的引入，增强了国家参考实验室的诊断能力。
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