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目的研究人类巨细胞病毒(human cytomegalovims,HCMV)UL144基因在先天性巨结肠(Hirschsorung’s disease,HD)临床株中的多态性,探讨HCMV UL144基因多态性与致病性之间的关系。方法随机选取53个先天性巨结肠患儿痉挛段结肠手术标本及经荧光定量PCR方法检测HC- MV DNA为阳性的4个HD患儿的尿标本,对照组为无症状或仅有皮肤轻度黄疸的6个尿标本。应用巢式聚合酶链反应的方法,扩增HCMVUL144基因开放阅读框架(ORF),扩增阳性的临床株进行双向 DNA测序,最后通过DNAclub、Bioedit、DNAstar、GeneDoc等软件进行分析。结果23份HD痉挛段肠组织(46％)及4份尿标本HCMVUL144基因扩增阳性,并且完成测序。种系进化树分析结果显示25个 HD患儿的DNA序列分为3个基因型,G1A型64．O％,G2型24％,G3型12％。与对照组比较,经X~2检验,X~2=10．93,p为0．012;其中HD临床株G1A和G3型基因经Fisher检验,P为0．015,差异具有统计学意义。全结肠型、长段型及普通型HD分散分布于UL144各个基因型中。结论HD与HCMV感染有关,HCMV可能是HD的病因之一;在HD患儿中,HCMV感染以UL144．基因G1A型为主;HD的临床分型与HCMVUL144基因分型无关。
Objective To study the polymorphism of human cytomegalovims (HCMV) UL144 gene in Hirschsorung’s disease (HD) clinical isolates and explore the relationship between HCMV UL144 gene polymorphism and pathogenicity. Methods Fifty-three Hirschsprung’s disease patients with sporadic colonic surgeries and 4 HD-positive children whose HC-MV DNA was positive were detected by fluorescence quantitative PCR. The control group was asymptomatic or had only mild skin Jaundice six urine samples. Nested polymerase chain reaction method was used to amplify HCMVUL144 open reading frame (ORF), amplified positive clinical strains for bidirectional DNA sequencing, and finally analyzed by DNAclub, Bioedit, DNAstar, GeneDoc and other software. Results The 23 HCSVUL144 genes in the HS spastic tract intestine (46%) and 4 urine specimens were positive for amplification and sequencing was completed. Phylogenetic tree analysis showed that DNA sequences of 25 HD patients were divided into three genotypes and G1A type 64. O%, G2 type 24%, G3 type 12%. Compared with the control group, X ~ 2 test showed that X ~ 2 = 10.93 and p = 0.012. Among them, the G1 and G3 genes of HD clinical strains were tested by Fisher test, P was 0.015, the difference was statistically significant. Colon-type, long-segment and general-type HD distributed in UL144 various genotypes. Conclusions HD is associated with HCMV infection and HCMV may be one of the causes of HD. In HD children, HCMV infection is associated with UL144. Gene G1A-based; HD clinical classification and HCMVUL144 genotyping nothing to do.