Kremer Jacques R, Nguyen Giang H, Shulga Sergey V, Nguyen Phuc H, Nguyen Ut T, Tikhonova Nina T, Muller Claude P
Institute of Immunology and WHO Collaborative Centre for Measles and WHO European Regional Reference Laboratory for Measles and Rubella, Laboratoire National de Santé, L-1011, Luxembourg.
J Med Virol. 2007 Jul;79(7):987-94. doi: 10.1002/jmv.20827.
The nucleoprotein genes of 49 measles virus (MV) strains circulating in Russia between 2000 and 2006 and in Vietnam in 2003 were analyzed by genotype-specific PCR and the results were compared with their sequences. The sequences revealed the presence of genotypes H1 and H2 in the center (Nha Trang) and the north (Hanoi) of Vietnam, respectively. The relative diversity of the H2 strains suggested an endemic circulation of these viruses in the capital. In contrast genotype H1 strains from Nha Trang were homogenous genetically, which may indicate a recent importation. The strains obtained from 12 different regions of the Russian Federation were assigned to the genotypes H1, D4, and D6. Most strains (81.4%) were correctly genotyped by a multiplex PCR method which was sensitive to genotype-specific mutations [Kremer et al. (2004): J Clin Microbiol 42: 3017-3022]. Ambiguous or negative results for some clade H and genotype D6 strains were due to point mutations in the type-specific primer binding sites. After exchanging a single nucleotide in both the clade H- and the genotype D6-specific primers, all strains were assigned correctly to their genotype. A simplified procedure for use in Vietnam was developed to distinguish directly between genotypes H1 and H2 and any non-H genotype. These results demonstrate that our multiplex PCR method can be adapted easily to new sequence variants or specific epidemiological situations, and thus be very useful for rapid genotyping of large number of samples even in laboratories which do not have sequencing facilities.
对2000年至2006年间在俄罗斯流行以及2003年在越南流行的49株麻疹病毒（MV）的核蛋白基因进行了基因型特异性PCR分析，并将结果与其序列进行了比较。序列分析显示，在越南中部（芽庄）和北部（河内）分别存在H1和H2基因型。H2毒株的相对多样性表明这些病毒在首都呈地方性流行。相比之下，来自芽庄的H1基因型毒株在基因上是同质的，这可能表明是近期输入的。从俄罗斯联邦12个不同地区获得的毒株被归类为H1、D4和D6基因型。大多数毒株（81.4%）通过对基因型特异性突变敏感的多重PCR方法被正确分型[克雷默等人（2004年）：《临床微生物学杂志》42：3017 - 3022]。一些H分支和D6基因型毒株的结果不明确或为阴性是由于型特异性引物结合位点的点突变。在H分支和D6基因型特异性引物中都交换一个核苷酸后，所有毒株都被正确地归类到其基因型。开发了一种适用于越南的简化程序，以直接区分H1和H2基因型以及任何非H基因型。这些结果表明，我们的多重PCR方法可以很容易地适应新的序列变异或特定的流行病学情况，因此即使在没有测序设施的实验室中，对于大量样本的快速基因分型也非常有用。
