Dries V, von Both I, Müller M, Gerken G, Schirmacher P, Odenthal M, Bartenschlager R, Drebber U, Meyer zum Büschenfeld K H, Dienes H P
Institute of Pathology, University of Cologne, Cologne, Germany.
Hepatology. 1999 Jan;29(1):223-9. doi: 10.1002/hep.510290118.
The diagnosis of hepatitis C is based on serological testing for antibodies against various epitopes of the hepatitis C virus (HCV) and detection of HCV RNA in serum, because anti-HCV antibodies alone cannot discriminate patients who are infectious from those who have resolved the infection. If HCV RNA is not detected, which is the case in at least 20% of enzyme immunoassay (EIA)-positive patients, diagnosis remains unclear in a state of disease possibly well suited for therapeutic intervention. Therefore, we investigated if detection of HCV antigens or HCV RNA in routinely processed, formalin-fixed and paraffin-embedded (ffpe) liver biopsy specimens of patients positive for anti-HCV, but negative for HCV RNA in serum, could confirm diagnosis in this serological constellation. We detected HCV RNA by reverse-transcription polymerase chain reaction (RT-PCR) in 27 (61%) of 44 ffpe liver biopsies from EIA-positive, but HCV-RNA-seronegative, patients. Testing of 18 of these biopsies by a panel of polyclonal antibodies against structural and nonstructural HCV proteins revealed positive immunostaining in 6 cases (33%), which were also positive by RT-PCR. Most biopsies showed necroinflammation compatible with chronic hepatitis C, and the detection of tissue HCV RNA correlated significantly with a higher grade of inflammatory activity. Detectability of HCV RNA did not correlate with HCV subtype. In conclusion, the search for HCV RNA by RT-PCR within the liver biopsy specimen can establish rapid and unequivocal diagnosis of hepatitis C in at least 60% of anti-HCV antibody-positive patients who are seronegative for HCV RNA, and thus may help to avoid repeated testing and delayed therapy. Tissue RT-PCR may also be more efficient than serological testing for surveillance of interferon therapy response, because ongoing chronic active hepatitis C is clearly demonstrated in the absence of detectable serum HCV RNA.
丙型肝炎的诊断基于对丙型肝炎病毒（HCV）各种表位的抗体进行血清学检测以及血清中HCV RNA的检测，因为仅抗-HCV抗体无法区分有传染性的患者和已清除感染的患者。如果未检测到HCV RNA（至少20%的酶免疫测定（EIA）阳性患者属于这种情况），则在可能非常适合进行治疗干预的疾病状态下，诊断仍不明确。因此，我们研究了在常规处理、福尔马林固定和石蜡包埋（ffpe）的抗-HCV阳性但血清HCV RNA阴性患者的肝活检标本中检测HCV抗原或HCV RNA，是否能在这种血清学情况下确诊。我们通过逆转录聚合酶链反应（RT-PCR）在44例EIA阳性但HCV-RNA血清阴性患者的ffpe肝活检标本中检测到27例（61%）有HCV RNA。用一组针对HCV结构和非结构蛋白的多克隆抗体对其中18例活检标本进行检测，发现6例（33%）免疫染色呈阳性，这些病例RT-PCR也呈阳性。大多数活检标本显示出与慢性丙型肝炎相符的坏死性炎症，并且组织HCV RNA的检测与更高等级的炎症活动显著相关。HCV RNA的可检测性与HCV亚型无关。总之，通过RT-PCR在肝活检标本中检测HCV RNA可在至少60%的抗-HCV抗体阳性但HCV RNA血清阴性的患者中快速明确诊断丙型肝炎，从而有助于避免重复检测和延迟治疗。对于监测干扰素治疗反应，组织RT-PCR可能也比血清学检测更有效，因为在未检测到血清HCV RNA的情况下可明确显示存在持续的慢性活动性丙型肝炎。
