CAJ | 학술논문

目的研究婴幼儿人巨细胞病毒(HCMV)临床分离株包膜糖蛋白H(gH)的基因多态性,分析该基因不同型别在HCMV感染相关的不同疾病中的分布情况及其与不同疾病之间的关系.方法收集2010年5月至2012年10月华中科技大学同济医学院附属同济医院血HCMV-IgM、IgG阳性的不同疾病住院患儿的新鲜尿标本进行病毒分离,应用巢式PCR扩增结合限制性片段长度多态性及测序对临床分离株gH基因进行分型.结果 102例临床分离株经巢式PCR扩增均为阳性(43例为婴儿肝炎综合征,38例为无黄疸型肝炎,13例为肺炎,7例为血小板减少性紫癜,1例为先天性CMV感染).102例均得到gH基因分型结果,其中gH1型62株,占60.8％；gH2型40株,占39.2％,以gH1型为主,未见混合型和发现新的型别.gH基因型别在婴儿肝炎综合征(43株分离株中26例为gH1)、无黄疸型肝炎(38株分离株中25例为gHl)及肺炎(13株分离株中9例为gH1)中与总体分布趋势一致(x2 =0.357,P＞0.05),而在血小板减少性紫癜中以gH2型别为主,7株分离株中6例为gH2(x2 =6.083,P＜0.05).结论本地区婴幼儿临床分离株gH基因型总体以gHl型占优势,在婴儿肝炎综合征、无黄疸型肝炎、肺炎等疾病中,gH基因型别分布与总体分布差异没有统计学意义,而在血小板减少性紫癜中以gH2型为主,提示gn型别可能与不同疾病表现存在一定的相关性.
목적 연구영유인인거세포병독(HCMV)림상분리주포막당단백H(gH)적기인다태성,분석해기인불동형별재HCMV감염상관적불동질병중적분포정황급기여불동질병지간적관계.방법 수집2010년5월지2012년10월화중과기대학동제의학원부속동제의원혈HCMV-IgM、IgG양성적불동질병주원환인적신선뇨표본진행병독분리,응용소식PCR확증결합한제성편단장도다태성급측서대림상분리주gH기인진행분형.결과 102례림상분리주경소식PCR확증균위양성(43례위영인간염종합정,38례위무황달형간염,13례위폐염,7례위혈소판감소성자전,1례위선천성CMV감염).102례균득도gH기인분형결과,기중gH1형62주,점60.8％；gH2형40주,점39.2％,이gH1형위주,미견혼합형화발현신적형별.gH기인형별재영인간염종합정(43주분리주중26례위gH1)、무황달형간염(38주분리주중25례위gHl)급폐염(13주분리주중9례위gH1)중여총체분포추세일치(x2 =0.357,P＞0.05),이재혈소판감소성자전중이gH2형별위주,7주분리주중6례위gH2(x2 =6.083,P＜0.05).결론 본지구영유인림상분리주gH기인형총체이gHl형점우세,재영인간염종합정、무황달형간염、폐염등질병중,gH기인형별분포여총체분포차이몰유통계학의의,이재혈소판감소성자전중이gH2형위주,제시gn형별가능여불동질병표현존재일정적상관성.
Objective To investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates,to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.Method Fresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive.Virus was isolated from these specimens.Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP),the purified PCR products were digested by restriction endonuclease HhaI.The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining.Classification and results of sequencing were compared.Result Totally 102 HCMV clinical isolates were obtained.Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome,38 cases had anicteric hepatitis,13 pneumonia,7 thrombocytopenic purpura,and 1 congenital CMV infection) were positive by nested-PCR,whose positive rate was 100％.The results showed that 62 strains were gH1 genotypes (60.8％),while 40 strains were gH2 (39.2％),mixed type or new genotype was not observed.In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes,17 clinical isolates were gH2 genotypes),anicteric hepatitis (25 were gH1,13 were gH2) and pneumonia (9 were gH1,4 were gH2),the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (x2 =0.357,P ＞0.05).However,the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2,1 were gH2,x2 =6.083,P ＜ 0.05).Conclusion Genotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants.There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome,anicteric hepatitis and pneumonia.However,gH2 was the dominant genotype in thrombocytopenic purpura.These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.