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本研究用Vero/Slam细胞从辽宁省2008～2011年流行性腮腺炎暴发和散发患者的临床标本中分离到13株流行性腮腺炎野病毒(Mumps virus,MuV),应用逆转录-聚合酶链反应(RT-PCR)针对MuV分离株的SH基因的316个核苷酸片段进行扩增,并对该产物进行序列测定。将这13株MuV与从GenBank下载的世界卫生组织(WHO)MuV基因型参考株一起进行分子流行病学研究。结果提示:除2011-015株外,辽宁省2008～2011年12株MuV分离株均属于F基因型,核苷酸和氨基酸同源性为94.9%～100%和83.3%～100%。与F基因型参考株序列相比,核苷酸和氨基酸同源性分别为92.4%～97.2%和96.5%～84.2%。表明2008～2011年辽宁省流行的F基因型MuV发生较大的型内变异。另外还发现F基因型MuV在SH基因上存在着特异性突变(CNt65,CNt105,G Nt137,C Nt192,C Nt239,GNT262),而其它基因型MuV在这些位点上均未发生改变。F基因型MuV在SH基因编码的氨基酸保守位点也发生变化。如:第2位上由S→P,第6位上由P→L,第23位上由T→N,第48位上由L→P/R。与基因分型有关的氨基酸三联体,2008-01-007毒株也发生了改变,由IML变为TMP。2011-015株病毒与F基因型参考株平均核苷酸和氨基酸同源性分别为87.5%和79.8%,与G型参考株平均核苷酸和氨基酸同源性分别为96.8%和97.4%,属于G基因型。该基因型为中国内地首次发现。
In this study, 13 Mumps virus (MuV) isolates were isolated from clinical samples of mumps outbreaks and sporadic cases from 2008 to 2011 in Liaoning Province using Vero / Slam cells. Reverse transcription polymerase chain reaction The reaction (RT-PCR) was performed on a 316 nucleotide fragment of the SH gene of MuV isolate and the product was sequenced. These 13 strains of MuV were molecularly epidemiologically studied together with the World Health Organization (WHO) MuV genotype reference strain downloaded from GenBank. The results suggest that all the 12 strains of MuV isolated from Liaoning Province in 2011-2011 belonged to F genotype except for 2011-015, with 94.9% -100% and 83.3% -100% nucleotide and amino acid homology. Compared with the F genotype reference strain, the nucleotide and amino acid homologies were 92.4% -97.2% and 96.5% -84.2%, respectively. The results showed that there was a large intra-type variation in the MuV genotype of F in Liaoning Province during 2008-2011. In addition, MuV was found to have specific mutations (CNt65, CNt105, GNT137, C Nt192, C Nt239, and GNT262) in the SH gene, while MuV in other genotypes did not change at these sites. The F genotype MuV also changes in the conserved sites of amino acids encoded by the SH gene. Such as: the second by the S → P, the sixth by the P → L, the 23rd by T → N, the 48th by L → P / R. Amino acid triplets associated with genotyping, 2008-01-007 The strain also changed from IML to TMP. The average nucleotide and amino acid homology of strains 2011-015 and F was 87.5% and 79.8%, respectively. The homology of nucleotide and amino acid with reference strains of type G was 96.8% and 97.4%, respectively. Belongs to the G genotype. The genotype was first discovered in Mainland China.