摘要
目的:探讨不孕女性中人巨细胞病毒(HCMV)感染流行株的包膜糖蛋白B(gB)基因型分布及其与不孕的相关性。方法:采用酶联免疫吸附试验(ELISA)检测300例女性不孕患者血清中HCMV-IgM抗体,ELISA阳性的患者采集晨尿接种于人胚肺成纤维细胞(HELF),提取细胞病变(CPE)阳性培养液中的病毒DNA,以巢式PCR(nest PCR)法扩增HCMVgB基因,利用限制性核酸内切酶HinfΙ、RsaΙ对HCMV gB基因进行限制性片段长度多态性(RFLP)分析判断基因型别;随机抽取11例送检测序,测序结果使用Clustal X软件与GenBank标准病毒株进行序列比对,用MEGA4.1构建核苷酸序列的基因树。结果:300例女性不孕患者中HCMV-IgM抗体阳性45例,阳性率为15.0%。45例患者晨尿接种HELF细胞,观察1月后检测到的病毒阳性37例,阳性率为12.3%。37例病毒阳性患者中最常见的基因型为gB 1型25例(67.6%),其次为gB 3型7例(18.9%)和gB 2型5例(13.5%),没有检测到gB4基因型。测序结果与GenBank标准病毒株HCMVADl69和Towne进行序列比对后,用MEGA4.1成功构建基因树。结论:女性不孕症的发生与HCMV感染有明显相关性,HCMV感染导致的不孕中最常见的gB基因型为gB 1型,其次是gB 3、gB 2型,未检测到gB 4型。
Objective： To explore the distribution of human cytomegalovirus （HCMV） envelope glycoprotein B （gB） genotypes in infertile human and its correlation with infertility. Methods ： ELISA was used to detect HCMV - IgM antibody in serum samples of 300 infertile women, the morning urine samples of ELISA positive patients were obtained to inoculate hmnan embryonic lung fibroblasts （HELF）, HCMV DNA was abstracted from cytopathic positive culture solution, nest PCR was used to amplify HCMV gB gene, restriction fragment length polymorphism （RFLP） was performed by restriction endonucleases Hinf I and Rsa I to analyze and determine genotypes; 11 cases were randomly selected to conduct gene sequencing, then the results were compared with GenBank standard strains by Clustal X software, MEGA 4. 1 software was used to construct gene tree of nucleotide sequences. Results： Among 300 infertile women, 45 women were found with positive HCMV- IgM antibody, the positive rate was 15. 0%. The morning urine samples of 45 patients were obtained to inoculate HELF, 37 patients were found with positive HCMV after one month, the positive rate was 12. 3%. Among 37 patients with positive HCMV, the most common genotype was gB1 genotype （25 patients, 67.6% ）, followed by gB3 genotype （7 patients, 18.9% ） and gB2 genotype （5 patients, 13.5% ） , and gB4 genotype was not found. After compared with GenBank standard strain HCMVAD 169 and Towne, gene tree was constructed by MEGA 4. 1 software. Conclusion： There is a significant correlation between the occurrence of female infertility and HCMV infection, among the infertile women induced by HCMV infection, the most common genotype is gB1 genotype, followed by gB3 genotype and gB2 genotype, and gB4 genotype is not found.
出处
《中国妇幼保健》
CAS
北大核心
2012年第32期5147-5150,共4页
Maternal and Child Health Care of China
基金
滨州医学院科技计划〔BY2010KJ003〕
关键词
不孕
巨细胞病毒
包膜糖蛋白B
基因型
Infertility
Cytomegalovirus
Envelope glycoprotein B
Genotype
