汉坦病毒基因分型方法的建立及其核苷酸序列特征分析

Genotyping and DNA sequence analysis of Hantaviruses

目的使用RTnestedPCR分型检测方法及核苷酸序列测定技术,对来源于福建省内HFRS监测点的鼠肺标本及病人血清进行基因分型并对部分标本的核苷酸序列进行分析比较。结果24份阳性鼠肺标本中检出率为95.8%;20份病人血清标本中仅11份扩增阳性,检出率分别为:83%(≤1周),12.5%(>1周)。扩增阳性标本中仅1份RTPCR分型为HTN型,其余均为SEO型,这与核苷酸序列分型结果相一致。结论近年福建省流行的汉坦病毒仍以SEO型为主。核酸序列比较分析发现,福建省内同一地区流行的SEO型汉坦病毒核苷酸的同源性很高,大于99%,而不同地区病毒间核酸序列变异比较大,尤其是永春和松溪这两个地区的毒株差异达到20%左右,可能是SEO型中两个新的亚型。

In this paper, the genotyping of Hantaviruses （HV） isolated from the lung tissues from infected Rattus norvegicus and sera of patients with hemorrhagic fever with renal syndrome （HFRS） in Fujian province is described. In addition, the DNA sequence analysis in part of cases are also included. The experimental results showed that the positive rate in 12 specimens from 24 HFRS Rattus norvegicus as detected with M gene and RT-nested PCR was 95.8%, hut only 11 specimens from 20 serum samples of patients with HFRS showed a positive rate of amplification of 83 % （ ≤one week） or 12.5 % （ 〉 one week） respectively. As demonstrated by RT-PCR genotyping, in the positive samples of amplification there was only one case showing the HTN genotype, the others all belonged to the SEO genotype. This result was just the same with that of nucleotide sequence typing. It is evident that the Hantaviruses isolated in the recent epidemic in Fujian province are mainly the SEO type. The nudeotide homology of the SEO type of Hantaviruses in the same area of Fujian province is rather high, accounting for more than 99 %, but there are large diversity between the different districts, especially in Yongchun and Songxi district, in which the identity of nucleotides is less than 20 %. It is possible that they are new subtypes of Seoul virus.