CAJ | 학술논문

目的：研究广州地区新生儿感染人巨细胞病毒（ HCMV）临床低传代分离株UL138基因序列特征与多态性。方法从10例被证实为临床HCMV感染的新生儿尿液标本中分离HCMV低传代临床病毒株，进行多重PCR鉴定。对临床分离株进行HCMV UL138基因扩增、克隆、鉴定、测序及呈报GenBank；同时在GenBank搜索HCMV UL138同源序列，行序列分析，应用生物信息学在线软件分析UL138基因翻译后修饰位点、二级结构及等电点。结果成功分离3株HCMV临床病毒株，分别命名为 HCMV D3、D2和 D52。克隆测序后呈报 GenBank 并被收录。收录序列号分别为：DQ180375、DQ180387和DQ180359。分析结果显示在HCMV临床低传代D3、D2和D52病毒株UL138基因全序列841个碱基中，存在16个位点变异，可能编码的蛋白质氨基酸残基总数为169个，其基因序列高度保守，核苷酸变异均为碱基替换，无插入或缺失，碱基替换导致了7处氨基酸的改变。HCMV UL138蛋白翻译后修饰位点在除Toledo株外的所有分离株中均高度保守；所有分离株UL138蛋白的预测等电点均为6．51。结论广州地区感染婴儿的HCMV UL138基因及其编码的氨基酸序列极为保守，但仍具有一定的多态性，这在HCMV临床感染与致病机制研究中具有重要意义。
목적：연구엄주지구신생인감염인거세포병독（ HCMV）림상저전대분리주UL138기인서렬특정여다태성。방법종10례피증실위림상HCMV감염적신생인뇨액표본중분리HCMV저전대림상병독주，진행다중PCR감정。대림상분리주진행HCMV UL138기인확증、극륭、감정、측서급정보GenBank；동시재GenBank수색HCMV UL138동원서렬，행서렬분석，응용생물신식학재선연건분석UL138기인번역후수식위점、이급결구급등전점。결과성공분리3주HCMV림상병독주，분별명명위 HCMV D3、D2화 D52。극륭측서후정보 GenBank 병피수록。수록서렬호분별위：DQ180375、DQ180387화DQ180359。분석결과현시재HCMV림상저전대D3、D2화D52병독주UL138기인전서렬841개감기중，존재16개위점변이，가능편마적단백질안기산잔기총수위169개，기기인서렬고도보수，핵감산변이균위감기체환，무삽입혹결실，감기체환도치료7처안기산적개변。HCMV UL138단백번역후수식위점재제Toledo주외적소유분리주중균고도보수；소유분리주UL138단백적예측등전점균위6．51。결론엄주지구감염영인적HCMV UL138기인급기편마적안기산서렬겁위보수，단잉구유일정적다태성，저재HCMV림상감염여치병궤제연구중구유중요의의。
Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.