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目的了解引起新生儿晚期黄疸患者人巨细胞病毒(HCMV)gB基因型的分布,探讨HCMVgB基因多态性与黄疸之间的关系。结论采用荧光定量PCR法检测本院新生儿科98例晚期黄疸新生儿标本HCMV-DNA含量,巢式聚合酶链反应扩增阳性标本HCMVgB基因,并进行DNA测序,绘制种系进化树。利用HinfⅠ和RsaⅠ对gB基因进行限制性片段长度多态性(RFLP)分析。结果 30例晚期黄疸新生儿HCMV荧光定量PCR检测阳性,阳性率为30.6%。种系进化树分析结果显示gB基因分为4个基因型,gB1型15株(50%),gB2型5株(16.7%),gB3型9株(30.0%),gB1/3混合型1株(3.3%)。以HCMV实验室标准株AD169作为参考,将序列进行同源性比较,gB1型同源性为94.7%～95.0%,gB2型同源性93.1%～93.4%,gB3型同源性94.7%,gB1/3型同源性93.7%。RFLP分析将gB基因分为4个基因型,分型结果与种系进化树分型一致。结论 HCMV感染是导致新生儿晚期黄疸的原因之一;gB基因的DNA序列比较保守,但仍存在一定的多态性,晚期黄疸新生儿中HCMV感染以gB1、gB3型为主。
Objective To investigate the genotype distribution of human cytomegalovirus (HCMV) gB in patients with neonatal jaundice and to explore the relationship between HCMVgB gene polymorphism and jaundice. Conclusion The HCMV-DNA content of 98 neonates with neonatal jaundice in our hospital was detected by real-time PCR. HCMVgB gene was amplified by nested polymerase chain reaction (PCR) and DNA sequencing was carried out to map the phylogenetic tree. The gB gene was analyzed by restriction fragment length polymorphism (RFLP) using Hinf Ⅰ and Rsa Ⅰ. Results 30 cases of neonatal jaundice HCMV fluorescence quantitative PCR positive, the positive rate was 30.6%. The results of phylogenetic tree analysis showed that gB gene was divided into 4 genotypes: 15 strains (50%) of gB1, 5 strains (gG3), 9 strains of gB3 (30.0%), 1 strain of gB1 / 3 (3.3%). Homology comparison of the sequence of HCMV laboratory standard strain AD169 showed that the homology of gB1 was 94.7% -95.0%, the homology of gB2 was 93.1% -93.4%, the homology of gB3 was 94.7%, gB1 / 3 homology 93.7%. RFLP analysis divided the gB genes into four genotypes, and the typing results were consistent with the phylogenetic tree classification. Conclusions HCMV infection is one of the causes of neonatal jaundice. The DNA sequence of gB gene is conservative, but there are still some polymorphisms. In neonates with advanced jaundice, HCMV infection is dominated by gB1 and gB3.