丙型肝炎患者抗病毒治疗后外周血单个核细胞内病毒 RNA 的检测及其与疗效的关系

Detection of hepatitis C virus RNA in the peripheral blood mononuclear cells in patients with chronic hepatitis C and its relationship with effect of anti-viral treatment

目的：观察慢性丙型肝炎患者在聚乙二醇干扰素α-2a （Peg IFNα-2a）联合利巴韦林（RBV）抗病毒治疗前、后血清及 PBMC 内 HCV RNA 的变化，初步探讨 PBMC 内 HCV RNA 检测的临床意义。方法2013年6月至2014年12月在广州市第八人民医院感染科就诊的慢性丙型肝炎患者20例，在治疗前、后的不同时间点（0、2、4、12、24、36、48周）采集患者外周血，分离血浆和 PBMC；采用精确荧光定量方法（Cobas TaqMan 实时 PCR）检测丙型肝炎患者血浆 HCV RNA；采用实时-PCR、套式PCR 两种方法分别检测 PBMC 内 HCV RNA。计数资料采用χ^2检验。结果 Cobas 精确定量检测各时间点血浆中 HCV RNA 分别为（5．92±0．86）、（2．77±1．32）、（1．30±1．72）、（0．15±0．47）、0、0和（0．06±0．26）lgIU／mL；各时间点差异有统计学意义（F ＝148．06，P ＜0．01）。12周时有18例患者血浆中 HCV RNA 完全清除。套式 PCR 较实时 PCR 阳性率高（均 P ＜0．01）；20例患者血浆病毒载量及PBMC 中 HCV RNA 检测结果发现，PBMC 中 HCV RNA 清除速率较血浆明显延迟，其中2例 PBMC中 HCV RNA 持续性存在并在治疗结束24周后复发。结论慢性丙型肝炎患者 PBMC 中可以检测到HCV RNA，且套式 PCR 检测较实时 PCR 检测阳性率高；抗病毒治疗对 PBMC 内、外的 HCV 均有效，但 PBMC 中 HCV RNA 清除速率较血浆明显延迟；干扰素抗病毒过程中 PBMC 内 HCV 清除缓慢，预示存在治疗后复发的风险。

Objective To detect the change of hepatitis C virus （HCV）RNA in the peripheral blood mononuclear cells （PBMC）and serum of patients with chronic hepatitis C （CHC）during treatment with peg-interferon α-2a （Peg IFNα-2a）plus ribavirin （RBV）,and to analyze the clinical significance of HCV RNA detection in PBMC.Methods The peripheral blood samples of 20 CHC patients who visited Department of Infectious Diseases in Guangzhou No.8 People′s Hospital from June 2013 to December 2014,were collected during treatment with Peg IFNα-2a＋RBV at different time points （week 0,2,4, 12,24,36 and 48）.Serum and PBMC were separated.Accurate fluorescence quantification assay （Cobas TaqMan real time polymerase chain reaction[PCR]）was used to detect HCV RNA level in serum,while real-time PCR and nest-PCR were applied to detect HCV RNA in PBMC.Categorical data were analyzed byχ^2 test.Results Accurate fluorescence quantification of serum HCV RNA showed that HCV RNA level decline rapidly after treatment （F = 148.06,P 〈 0.01 ）,and 18 patients achieved HCV RNA undetectable at week 12 of treatment.The positive rate of nest-PCR was higher than real-time PCR （all P 〈0.01）.Comparison of HCV RNA levels in serum and PBMC from 20 cases found that,the clearance rate of HCV RNA in PBMC was postponed.Two patients whose HCV RNA in PBMC kept detectable＆amp;nbsp;relapsed at week 24 after end of treatment.Conclusions HCV RNA can be detected in PBMC of CHC patients and the positive rate of nest-PCR is higher than real-time PCR.Antiviral therapy is effective on HCV both inside and outside PBMC,but the clearance rate of HCV RNA in PBMC is postponed compared with that in serum.Slow clearance of HCV in PBMC may be a risk factor for relapse after end of treatment.