原位杂交在生精细胞巨细胞病毒感染检测中的应用

目的探讨男性不育患者精液中生精细胞人类巨细胞病毒（HCMV）感染状况，评估原位杂交技术（ISH）在生精细胞HCMVDNA检测的价值。方法收集119例精液检查中发现较多生精细胞男性不育病例精液，洗涤收集细胞后提取DNA，然后用聚合酶链反应（PCR）行HCMV筛查，阳性者精液细胞涂片用ISH行HCMVDNA检测。结果119例精液样本PCR检测HCMV阳性12例。12例HCMV阳性病例精液细胞涂片ISH检测生精细胞HCMV DNA均见阳性表达。HCMV阳性病例精液均发现较多未成熟生精细胞，并有不同程度的病理性损害。精子密度HCMV阳性组显著低于阴性组（P〈0．05）。HCMV阳性患者血清HCMV-IgG均阳性，HCMV-IgM均阴性。结论生精细胞HCMV感染时可出现不同程度的病理性损害，影响生精功能，可能是引起男性不育的原因之一；ISH可作为生精细胞HCMV感染的一种较理想联合检查方法。

The application of in stitu hybridization on detection of human cytomegalovirus in spermatogenic cells

Objective To investigate the infection status of human cytomegalovirus （HCMV） of spermatogenic cells in the semen of infertile male, in order to value the technology of in stitu hybridization（ISH）used in detection of HCMV DNA in spennatogenic cells.Methods 119 semen samples of infertile male were collected which more spermatogenic cells were found through semen routine assaying. Then washed and concentrated cells, and extracted DNA, and then detected HCMV DNA by polymerase chain reaction （PCR）. ISH technique was used to detect the expressions of HCMV DNA of the positive infected semen cells. Results 12 results of HCMV PCR were positive in 119 semen samples. Among all 12 HCMV PCR positive cases, HCMV DNA was positive expression in spermatogenic ceils by ISH. Among all HCMV positive cases, more immature spermatogenic cells were found in semen by microscope, and could find some variational pathologic alternation. Sperm density of HCMV positive group were significantly lower than that of HCMV negative group（ P 〈 0.05）.Among all HCMV positive cases, the HCMV-IgG was positive in blood and the HCMV-IgM was negative. Conclusion Spermatogenic cells with HCMV infection may cause pathologic lesions, and affect spermatogenesis. And these lesions may be one causational factors for male infertility. ISH is an ideal cooperative method to exam the infection of the spermatogenic cells.