摘要
目的对一例疑似麻疹病例进行实验室诊断,同时对病毒种系进化进行分析。方法应用Measles IgM试剂盒(μ-capture)对疑似患者血清进行病毒IgM抗体的检测,采集咽拭子标本,提取总RNA,合成麻疹病毒核蛋白基因特异性扩增引物,应用RT-PCR及Real-time PCR进行扩增,回收阳性扩增产物,进行序列测定,将所测序列与国内外麻疹病毒株序列进行核苷酸同源性分析,应用MEGA5.0软件构建核蛋白基因种系进化树。结果疑似病例血清麻疹病毒IgM抗体为阳性;RT-PCR法扩增病毒核蛋白基因可见一约330bp大小的条带;与Genbank报道的国内Mvi Hunanstrain 1、MviHunan strain 2、MviHunan strain 3的同源性最高;种系进化分析表明MviHunan Yueyang strain同所有H1型的麻疹病毒聚集在进化树的B组内,为麻疹病毒H型H1亚型。结论分离的病毒为麻疹病毒H1型,基因分型技术能进一步对病毒亚型进行鉴定。
Objective To make laboratory diagnosis of a suspected measles patient and to make a phylogenetic analysis on the measles virus.Methods The IgM antibody of measles virus in the suspected patient was detected by measles IgM diagnose kit（μ-capture method）.Total RNA was extracted from the throat swab sample before the nucleoprotein gene of the measles virus was amplified by RT-PCR and Real-time PCR with specific primers designed.The PCR positive product was recycled and purified before being sequenced.The nucleotide homology was analyzed by comparing the obtained sequence with the measles virus sequences from the Genbank.The phylogenetic tree was constructed by MEGA5.0 software.Results The IgM antibody of the measles virus in the suspected patient was positive.The PCR product of the nucleoprotein gene was a band about 330 bp,which had a high homology with the Mvi Hunan strain 1,Mvi Hunan strain 2 and Mvi Hunan strain 3 reported in the Genbank.Phylogenetic analysis showed that the isolated Mvi Hunan Yueyang strain and all H1 genotype of measles viruses clustered in the B group of the phylogenetic tree.It belonged to the H1 genotype of measles virus.Conclusions The isolated measles virus in the suspected measles patient belongs to H1 genotype.Genotyping technique can further identify the subtype of the virus.
出处
《实用预防医学》
CAS
2012年第12期1886-1888,共3页
Practical Preventive Medicine
关键词
麻疹
麻疹病毒
核蛋白基因
分型
种系进化
Measles
Measles virus
Nucleoprotein gene
Genotyping
Phylogenetic
