Barkholt L M, Johansson B, Veress B, Andersson J P, Ehrnst A
Department of Transplantation Surgery, Huddinge Hospital, Karolinska Institute, S-14186 Huddinge, Sweden.
Clin Diagn Virol. 1995 Aug;4(2):121-34. doi: 10.1016/0928-0197(95)00002-p.
In liver transplant (LTX) patients, cytomegalovirus (CMV) hepatitis as a cause of graft dysfunction occurs in 15-25% of the patients. Polymerase chain reaction (PCR), applied to liver biopsy specimens, may increase the ability to detect CMV DNA at a local site. In this study, PCR was used to compare its relation to the development of clinical CMV hepatitis.
Nested polymerase chain reaction (nPCR), derived from a conserved region of the CMV major immediate-early gene, was used to examine 141 frozen liver biopsies from 61 LTX patients for the presence of CMV DNA. 134 biopsies were obtained from 54 patients with pathological liver function tests within four months after transplantation. The remaining seven patient biopsies were derived from the one-year investigation after LTX and served as controls. The results were compared to virus isolation, antigen detection by immunohistology and in situ hybridization for CMV DNA of the biopsy specimens. Histological examination was performed to verify a diagnosis of viral hepatitis.
CMV DNA was amplified in 11% (15/134) of the biopsies, corresponding to 20% (11/54) of the patients. Virus isolation revealed CMV in 5% (7/134) of the samples. None of the nPCR-negative biopsies was virus culture positive. CMV genomes were detected by nPCR more frequently than CMV hepatitis was diagnosed by using the combination of CMV-specific histopathology and/or immunohistology and/or CMV-positive virus isolation (p < 0.01). However, when this comparison was performed within individual patients, the difference was not significant (p > 0.05). If the results of in situ hybridization were included in the diagnostic criteria of CMV hepatitis, the nPCR was comparable to these, both at the biopsy and the patient levels (p > 0.1 and p > 0.05, respectively). For the diagnosis of CMV hepatitis the negative predictive value of CMV-nPCR was 1.0. The positive predictive value ranged from 0.55 to 0.82 depending on the criteria of CMV hepatitis. The nPCR also detected signs of CMV infection in the liver graft earlier than virus isolation, 11 versus 21 days, respectively, after transplantation.
The frequency of CMV DNA positivity, measured by nPCR, was similar to that revealed by other combined methods. We suggest that the combined findings of histological cholangitis and/or lobulitis together with nPCR for CMV DNA can be used as a diagnostic criterion for initiation of antiviral treatment against CMV hepatitis.
在肝移植（LTX）患者中，巨细胞病毒（CMV）性肝炎作为移植物功能障碍的一个病因，在15% - 25%的患者中出现。应用于肝活检标本的聚合酶链反应（PCR），可能会提高在局部部位检测CMV DNA的能力。在本研究中，使用PCR来比较其与临床CMV性肝炎发生发展的关系。
源自CMV主要立即早期基因保守区的巢式聚合酶链反应（nPCR），用于检测61例LTX患者的141份冷冻肝活检标本中CMV DNA的存在情况。134份活检标本取自54例移植后4个月内肝功能检查异常的患者。其余7例患者的活检标本取自LTX术后1年的调查，作为对照。将结果与病毒分离、免疫组织化学抗原检测以及活检标本CMV DNA的原位杂交结果进行比较。进行组织学检查以核实病毒性肝炎的诊断。
11%（15/134）的活检标本中扩增出CMV DNA，对应20%（11/54）的患者。病毒分离显示5%（7/134）的样本中有CMV。nPCR阴性的活检标本中无一例病毒培养阳性。通过nPCR检测到CMV基因组的频率高于通过CMV特异性组织病理学和/或免疫组织化学和/或CMV阳性病毒分离联合诊断CMV性肝炎的频率（p < 0.01）。然而，在个体患者中进行这种比较时，差异不显著（p > 0.05）。如果将原位杂交结果纳入CMV性肝炎的诊断标准，nPCR在活检和患者水平上与之相当（分别为p > 0.1和p > 0.05）。对于CMV性肝炎的诊断，CMV - nPCR的阴性预测值为1.0。阳性预测值根据CMV性肝炎的标准在0.55至0.82之间。nPCR在肝移植中检测到CMV感染迹象也比病毒分离更早，分别在移植后11天和21天。
通过nPCR检测到的CMV DNA阳性频率与其他联合方法显示的频率相似。我们建议，组织学胆管炎和/或小叶炎的联合发现以及CMV DNA的nPCR检测结果可作为启动抗CMV性肝炎抗病毒治疗的诊断标准。
