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《Chinese Journal of Microbiology and Immunology》 2003-01
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A study on variability of human cytomegalovirus UL144 gene in low-passage clinical isolates

HE Rong, RUAN Qiang, JI Yao-hua, LIU Qing, CHEN Shu-rong, LIU Lan-qing, L Sheng-min. Virus Laboratory, 2nd Clinical Hospital, China Medical University, Shenyang 110004, China Corresponding author: HE Rong, E-mail: keren -rong@sina.com  
Objective Human cytomegalovirus (HCMV) is an important infectious factor that results in neonatal disease and congenital deformity. HCMV may invade many organs. The different symptoms and tissue tropism of HCMV infection perhaps result from the genetic polymorphism of HCMV. Recent study showed that Toledo genome contained 19 open reading frames (denoted UL131 to 151) which were not present in the AD169 genome, leading us to focus on the relationship between HCMV disease and the products of these 19 open reading frames. UL144 open reading frames encode a homologue of the tumor necrosis factor receptor. It seems important to study the strain-specific variability of UL144 sequence in low-passage clinical isolates and to discuss if the variability related to the clinical HCMV infection. Methods HCMV-UL144 gene was amplified by PCR assay in 65 low-passage clinical isolates and urine from 7 healthy children, which were HCMV-DNA positive as shown by QPCR. All the positive PCR products were analyzed by HMA-SSCP (Heteroduplex mobility assay, single-stranded conformation polymorphism) and 32 of them were sequenced. The UL144 sequences of 32 clinical isolates have been assigned GenBank accession No. AF382014 to AF382031 and AF447377 to AF447390. Results 55 isolates and 5 urine samples were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in UL144 ORFs. Phylogenetic analysis indicated that the nuclear sequences could be classified into 3 major genotypes. Comparing between UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. Genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively. Conclusion HCMV-UL144 exist in almost all the low passage isolates and the sequence is hypervariability. The characters of sequences in different kinds of isolates showed that UL144 gene might play an important role in HCMV infection. [
【Fund】: 国家自然科学基金资助项目 ( 30 170 986 )
【CateGory Index】: R373
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