Sarrazin C, Hendricks D A, Sedarati F, Zeuzem S
Medizinische Klinik II, Johann Wolfgang Goethe-Universität, 60590 Frankfurt am Main, Germany.
J Clin Microbiol. 2001 Aug;39(8):2850-5. doi: 10.1128/JCM.39.8.2850-2855.2001.
Transcription-mediated amplification (TMA) is an isothermal, autocatalytic target amplification method which has the potential to detect less than 50 hepatitis C virus (HCV) RNA copies/ml (10 IU/ml). The TMA assay was used to assess the presence of residual HCV RNA in plasma from patients treated with polyethylene glycol-modified interferon alpha-2a (peginterferon alpha-2a) who showed a virologic relapse after the end of therapy. Stored end-of-treatment and end-of-follow-up plasma samples from 177 of 267 patients treated with peginterferon alpha-2a (S. Zeuzem et al., N. Engl. J. Med. 343:1666--1672, 2000) were available for retesting by TMA. Plasma samples from patients in the same study who exhibited virologic relapse after treatment with standard interferon alpha-2a served as controls. Virologic response during the trial was defined as HCV RNA that was undetectable using a PCR-based test system with a sensitivity of 50 IU/mL (Cobas Amplicor HCV version 2.0) and was compared with TMA-based retesting results (VERSANT HCV RNA Qualitative Assay). Residual HCV RNA was detected in 4 of 60 cases (7%) by the TMA technology in end-of-treatment plasma samples from patients who relapsed after receiving peginterferon alpha-2a and in 6 of 18 patients (33%) following therapy with standard interferon alpha-2a. For peginterferon alpha-2a-treated patients with sustained virologic response, HCV RNA was detectable by TMA in end-of-treatment samples in 3 of 78 cases but in none of the end-of-follow-up samples. For all end-of-treatment and end-of-follow-up plasma samples of virologic nonresponders, a complete concordance between the PCR-based assay and TMA was observed. In conclusion, in patients with virologic relapse after the end of therapy, according to PCR, who were treated with peginterferon alpha-2a or standard interferon alpha-2a, residual HCV RNA was detectable in end-of-treatment samples by the TMA-based assay in 7 or 33% of cases, respectively. The lower rate of residual HCV RNA detection by TMA for patients treated with peginterferon alpha-2a than that for patients treated with standard interferon alpha-2a may be due to the maintained antiviral pressure of the long-acting peginterferon alpha-2a at the end-of-treatment visit.
转录介导扩增(TMA)是一种等温、自催化的靶标扩增方法,它有潜力检测出每毫升少于50个丙型肝炎病毒(HCV)RNA拷贝(10国际单位/毫升)。TMA分析用于评估接受聚乙二醇修饰的干扰素α-2a(聚乙二醇化干扰素α-2a)治疗后出现病毒学复发的患者血浆中残留HCV RNA的存在情况。对接受聚乙二醇化干扰素α-2a治疗的267例患者中的177例(S. Zeuzem等人,《新英格兰医学杂志》343:1666 - 1672, 2000)储存的治疗结束和随访结束时的血浆样本,可用TMA进行重新检测。同一研究中接受标准干扰素α-2a治疗后出现病毒学复发的患者的血浆样本用作对照。试验期间的病毒学应答定义为使用灵敏度为50国际单位/毫升的基于PCR的检测系统(Cobas Amplicor HCV版本2.0)检测不到HCV RNA,并与基于TMA的重新检测结果(VERSANT HCV RNA定性分析)进行比较。在接受聚乙二醇化干扰素α-2a治疗后复发的患者的治疗结束时血浆样本中,通过TMA技术在60例中的4例(7%)检测到残留HCV RNA;在接受标准干扰素α-2a治疗的18例患者中的6例(33%)检测到残留HCV RNA。对于聚乙二醇化干扰素α-2a治疗后获得持续病毒学应答的患者,在78例中的3例治疗结束时样本中通过TMA可检测到HCV RNA,但随访结束时样本中均未检测到。对于病毒学无应答者的所有治疗结束和随访结束时的血浆样本,基于PCR的检测和TMA之间观察到完全一致。总之,在治疗结束后根据PCR出现病毒学复发且接受聚乙二醇化干扰素α-2a或标准干扰素α-2a治疗的患者中,基于TMA的检测分别在7%或33%的治疗结束时样本中检测到残留HCV RNA。聚乙二醇化干扰素α-2a治疗患者通过TMA检测到残留HCV RNA的比例低于标准干扰素α-2a治疗患者,这可能是由于长效聚乙二醇化干扰素α-2a在治疗结束时维持了抗病毒压力。