[无症状感染儿童临床分离HCMV UL144基因多态性分析]。

Gangqiang Guo, Shangdan Xie, Sisi Ye, Liang Zhang, Xiangwei Sun, Baoqing Li, Lifang Zhang, Xiangyang Xue
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摘要

本研究的目的是检测人类巨细胞病毒(HCMV) ul144基因在无症状HCMV感染儿童中的多态性。采用PCR方法扩增无症状HCMV-DNA阳性儿童尿液标本中的UL144开放阅读框(ORF),并对两条扩增子链进行测序。使用BioEdit、DNAstar、mega5.0、GeneDoc等软件进行序列分析。50株临床菌株中有21株扩增成功并测序,阳性率为42%。核苷酸序列同源性为80.2% ~ 100%,氨基酸序列同源性为77.8% ~ 100%。UL144序列分布在A型(47.61%)和B型(52.38%)两个基因型中。使用Expasy数据库分析UL144蛋白的重要功能基序。结果表明,与a型相比,B型UL144在1 ~ 16个氨基酸残基之间和30 ~ 96个氨基酸残基之间分别增加了一个proka - lipoprotein位点和一个ZF-CTCHY位点。与Toledo菌株的UL144基因相比,a型UL144在CRD1和crd2上具有较高的保守性。而在UL144 B型的CRD1和crd2中观察到明显更多的变异。在UL144 A型和B型中,跨膜和细胞质结构域高度保守。UL144 A型和B型核苷酸序列的变化不会导致UL144蛋白的预测等电点或二级结构发生重大变化。无症状HCMV感染儿童UL144基因型分为A型和B型,与有症状HCMV感染儿童不同。
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[Analysis of Polymorphisms in the HCMV UL144 Gene in Clinical Isolates from Children with Asymptomatic Infection].

The purpose of this study was to examine polymorphisms in the human cytomegalovirus(HCMV)UL144gene in children with asymptomatic HCMV infection. PCR was performed to amplify the UL144 open reading frame(ORF)from urine specimens of asymptomatic HCMV-DNA positive children and both strands of the amplicon were sequenced. Sequence analysis was performed with software including BioEdit,DNAstar,Mega5.0and GeneDoc. Twenty-one of 50 clinical strains were successfully amplified and sequenced, giving a positive rate of detection of 42%.Nucleotide sequence homology ranged from 80.2%to100% and amino acid sequence homology ranged from 77.8%to 100%.The UL144 sequences were distributed among two genotypes, type A(47.61%)and type B(52.38%).The Expasy database was used to analyze the important functional motifs of the UL144 protein. These results revealed that there was a high level of conservation of post-translational modification sites including ASN, PKC, TNFR, and NCD3 G.UL144type B added a PROKAR-LIPOPROTEIN site and ZF-CTCHY site between amino acid residues 1 and 16 and between amino acid residues 30 and 96,respectively,as compared with type A.Compared to the UL144 gene from the Toledo strain, there was a high level of conservation in the CRD1 and CRD2of UL144 type A, while significantly more variability was observed in CRD1 and CRD2of UL144 type B. The transmembrane and cytoplasmic domains were highly conserved in both UL144 type A and type B. Variation in nucleotide sequences of UL144 type A and type B did not cause major changes to the predicted isoelectric point or secondary structure of the UL144 protein. The UL144 genotype of children with asymptomatic HCMV infection was divided into type A and type B, which was different from children with symptomatic HCMV infection.

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