An Le Van, Bao Chi Le Thi, Bach Nguyen Hoang, Hai Duong Huynh Thi, Deligios Massimo, Alberti Alberto, Cappuccinelli Piero
Department of Microbiology, Hue University of Medicine and Pharmacy, Thua Thien Hue, Vietnam.
J Infect Dev Ctries. 2013 Mar 14;7(3):235-42. doi: 10.3855/jidc.2883.
The influenza A(H1N1)pdm09 virus arrived in Vietnam in May 2009 via the United States and rapidly spread throughout the country. This study provides data on the viral diagnosis and molecular epidemiology of influenza A(H1N1)pdm09 virus isolated in Thua Thien Hue Province, central Vietnam.
Nasopharyngeal swabs and throat swabs from 53 clinically infected patients in the peak of the outbreak were processed for viral diagnosis by culture and RT-PCR. Sequencing of entire HA and NA genes of representative isolates and molecular epidemiological analysis were performed.
A total of 32 patients were positive for influenza A virus by virus culture and/or RT-PCR; of these 22 were positive both by viral isolation and RT-PCR, 2 only by virus culture and 8 only by RT-PCR. The novel subtype of influenza A(H1N1)pdm09 was present in 93.4% of the isolates. Phylogenetic analysis of the HA and NA gene sequences showed identities higher than 99.50% in both genes. They were also similar to reference isolates in HA sequences (>99% identity) and in NA sequences (>98.50% identity). Amino acid sequences predicted for the HA gene were highly identical to reference strains. The NA amino acid substitutions identified did not include the oseltamivir-resistant H275Y substitution.
viral isolation and RT-PCR together were useful for diagnosis of the influenza A(H1N1)pdm09 virus. Variations in HA and NA sequences are similar to those identified in worldwide reference isolates and no drug resistance was found.
甲型H1N1pdm09流感病毒于2009年5月经美国传入越南,并迅速在全国传播。本研究提供了越南中部顺化省分离出的甲型H1N1pdm09流感病毒的病毒诊断和分子流行病学数据。
对疫情高峰期53例临床感染患者的鼻咽拭子和咽拭子进行培养和逆转录聚合酶链反应(RT-PCR)以进行病毒诊断。对代表性分离株的整个血凝素(HA)和神经氨酸酶(NA)基因进行测序并进行分子流行病学分析。
通过病毒培养和/或RT-PCR,共有32例患者甲型流感病毒呈阳性;其中22例通过病毒分离和RT-PCR均呈阳性,2例仅通过病毒培养呈阳性,8例仅通过RT-PCR呈阳性。93.4%的分离株为新型甲型H1N1pdm09亚型。HA和NA基因序列的系统发育分析显示,两个基因的同源性均高于99.50%。它们在HA序列(同源性>99%)和NA序列(同源性>98.50%)上也与参考分离株相似。预测的HA基因氨基酸序列与参考毒株高度相同。鉴定出的NA氨基酸替代不包括对奥司他韦耐药的H275Y替代。
病毒分离和RT-PCR共同用于甲型H1N1pdm09流感病毒的诊断。HA和NA序列的变异与全球参考分离株中鉴定出的变异相似,未发现耐药性。