Department of Virology, Tehran University of Medical Sciences, Tehran, Iran.
J Med Virol. 2011 Jun;83(6):989-95. doi: 10.1002/jmv.22044.
The diagnosis of cryptogenic liver disease is made when after extensive evaluations, recognizable etiologies of chronic liver disease are excluded. In this study, the presence of hepatitis C virus (HCV) RNA was tested in peripheral blood mononuclear cells (PBMCs) taken from Iranian patients who although were found negative for plasma HCV RNA and anti-HCV antibodies, suffered from chronic liver disease of unknown etiology. From September 2007 to March 2010, 69 patients from Tehran with chronic liver disease of unknown etiology who were referred to our center were enrolled in the present study. PBMCs were isolated from 10 mL peripheral blood specimens. HCV-RNA status was tested in plasma and PBMCs samples by reverse-transcription polymerase chain reaction (RT-PCR). HCV-RNA was detected in HCV-positive PBMCs specimens by RT-PCR method. HCV genotypes were subsequently analyzed in HCV-positive samples using restriction fragment length polymorphism (RFLP) assay; then HCV genotypes were confirmed by sequencing of 5' non-coding fragments after cloning PCR products into pJET1.2/blunt cloning vector. HCV-RNA was detected in PBMCs specimens belonging to 7 (10%) out of 69 patients. Genotyping of the HCV-RNA isolated from PBMCs showed that 3 (43%) patients with occult HCV infection had genotype 1b, 2 (29%) had genotype 1a, and another 2 (29%) had genotype 3a. The results of this study suggest that patients with chronic liver disease of unknown etiology may have occult HCV infection in the absence of anti-HCV antibodies and plasma HCV-RNA. It has been suggested that in the absence of liver biopsy specimens, analysis of PBMC sample for HCV-RNA would be informative.
当在广泛评估后排除了慢性肝病的可识别病因时，即可诊断为隐匿性肝病。在本研究中，对来自伊朗的慢性病因不明的肝病患者的外周血单核细胞 (PBMC) 进行了检测，尽管这些患者的血浆 HCV RNA 和抗 HCV 抗体均为阴性，但仍存在 HCV RNA。2007 年 9 月至 2010 年 3 月，本中心收治了 69 例来自德黑兰的慢性病因不明的肝病患者，纳入本研究。从 10mL 外周血标本中分离 PBMC。采用逆转录聚合酶链反应 (RT-PCR) 检测血浆和 PBMC 样本中的 HCV-RNA 状态。采用 RT-PCR 法检测 HCV 阳性 PBMC 标本中的 HCV-RNA。随后在 HCV 阳性样本中使用限制性片段长度多态性 (RFLP) 分析 HCV 基因型；然后通过克隆 PCR 产物到 pJET1.2/ blunt 克隆载体后对 5'非编码片段进行测序来确认 HCV 基因型。在 69 例患者中，有 7 例（10%）患者的 PBMC 标本中检测到 HCV-RNA。从 PBMC 中分离出的 HCV-RNA 的基因分型显示，3 例（43%）隐匿性 HCV 感染患者为 1b 基因型，2 例（29%）为 1a 基因型，另 2 例（29%）为 3a 基因型。本研究结果提示，在缺乏抗 HCV 抗体和血浆 HCV-RNA 的情况下，慢性病因不明的肝病患者可能存在隐匿性 HCV 感染。有人建议，在缺乏肝活检标本的情况下，分析 PBMC 样本中的 HCV-RNA 会提供有价值的信息。
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