组织学、血清学、尿荧光定量聚合酶链反应及尿沉渣中巨细胞包涵体检测方法在诊断巨细胞病毒所致胆汁淤积型婴儿肝炎综合征作用的研究

Study of histology,serology,urine HCMV-DNA-QPCR and on diagnosis of infantile cytomegalovirus hepatitis with cholestasis

目的:探讨肝脏光镜组织学检查、血清学检测巨细胞病毒(Human cytomegalovirus,HCMV)特异性免疫球蛋白M(Immune globulin M,IgM)、尿HCMV荧光定量PCR(Fluorescence quantitative polumerase chain reaction,QPCR)检测和晨尿找包涵体方法在诊断婴儿肝炎综合征中HCMV感染的作用。方法:免疫组织化学法检测肝组织HCMV的极早期抗原(Immediatelyearlyantigen,IEA)和早期抗原(Early antigen,EA)。采用Fisher精确检验比较HCMV-IEA/EA阳性组和阴性组在组织形态改变上的差异。运用ELISA方法常规检测血清HCMV抗体IgM、荧光定量PCR技术检测尿HCMV-DNA和瑞氏染色后显微镜下检测晨尿脱落细胞沉渣中包涵体,分析上述检查方法的灵敏度和特异度。结果:淤胆型婴儿肝炎综合征(Infantilehepatitissyndrome,IHS)患儿肝组织HCMV-IEA/EA阳性22/36例。HCMV-IEA/EA阳性组及阴性组比较,光镜组织形态学分析无明显特异性,晨尿脱落细胞找HCMV包涵体差异无统计学意义(P均>0.05),而血清HCMV-IgM、荧光定量PCR技术检测尿HCMV-DNA差异有统计学意义(P均<0.01)。血清HCMV-IgM、荧光定量PCR技术检测尿HCMV-DNA和晨尿脱落细胞找HCMV包涵体诊断符合率分别为77.8%、69.4%和50.0%。结论:普通组织病理对淤胆型婴儿HCMV肝炎的诊断缺乏诊断特异性,血清HCMV-IgM检测仍不失为简单易行的实验室诊断方法。

Objective：To evaluate the feasibility and utility of Pathologic changes of liver biopsy,HCMV-IgM,urine HCMV-DNA-QPCR and inclusion detected in morning urine exfoliative cytes on diagnosis of cytomegalovirus infection in infantile hepatitis syndrome.Methods：Immediately early antigen（IEA）and early antigen（EA）of HCMV were detected in liver tissues by immunohistochemistry stain assay.Pathologic changes were compared between HCMV antigen positive samples and negative samples,and analyzed statistically by fisher exact test.Serum HCMV-IgM was detected by ELISA,urine HCMV-DNA was detected by real-time quantitative PCR assay and inclusion was detected in morning urine samples with Wright＇s staining under microscope.Specificities and sensitivities were evaluated.Results：HCMV-IEA/EA was positive in liver tissues of 22/36 cases.There was no significant difference in pathologic changes between HCMV-IEA/EA positive group and negative group under light microscope analyzed by SPSS software.There were significant differences for serum HCMV-IgM detection and urine HCMV-DNA real-time quantitative PCR assay（P〈0.01）and there was no significant difference for morning urine HCMV inclusion detection（P〉0.05）in HCMV-IEA/EA positive group and negative group.In HCMV-IEA/EA positive group,the specificity of serum HCMV-IgM detection,urine HCMV-DNA real-time quantitative PCR assay detection or morning urine HCMV inclusion detection was 71.4%,100% or 78.6% respectively.And the sensitivity was 81.8%,50.0% or 31.8% respectively.Conclusion：There are no specific pathologic changes in HCMV hepatitis in infants.Morning urine HCMV inclusion detection is not a specific method for the HCMV infection.Serum HCMV-IgM detection is still an expedient and utility method clinically.