未治疗和治疗慢性患者的标准化乙型肝炎病毒 RNA 定量：感染

StephenW

未治疗和治疗慢性患者的标准化乙型肝炎病毒 RNA 定量：感染随访的有希望的标志
Maria Francesca Cortese 1 2 3，Mar Riveiro-Barciela 3 4，David Tabernero 2 3，Francisco Rodriguez-Algarra 5，Adriana Palom 4，Sara Sopena 2，Ariadna Rando-Segura 2 6，Luisa Roade 4，Alison Kuchta 7，Roser Ferrer -Costa 8，Josep Quer 3 9，Beatriz Pacin 1 2 3，Marta Vila 1 2，Rosario Casillas 1 2，Selene Garcia-Garcia 1 2 3，Rafael Esteban 3 4，Tomás Pumarola 6 10，Maria Buti #3 4，Francisco罗德里格斯-弗里亚斯 # 1 2 3
隶属关系
隶属关系

    1
    临床生物化学，Vall D'hebron 研究所，巴塞罗那自治大学，巴塞罗那，西班牙。
    2
    生物化学和微生物学，肝脏病理学部门，Vall D'hebron 大学医院，巴塞罗那自治大学，巴塞罗那，西班牙。
    3
    Centro De Investigación Biomédica En Red，Enfermedades Hepáticas y Digestvas (CIBERehd)，Instituto De Salud Carlos III，马德里，西班牙。
    4
    西班牙巴塞罗那自治大学 Vall D'hebron 大学医院内科肝脏科。
    5
    暴雪研究所、Barts 和伦敦玛丽女王大学伦敦医学和牙科学院，英国伦敦。
    6
    病毒学部门，微生物学部，Vall D'hebron 大学医院，巴塞罗那自治大学，西班牙巴塞罗那。
    7
    罗氏分子系统公司，美国加利福尼亚州普莱森顿。
    8
    西班牙巴塞罗那瓦尔德希伯伦大学医院生物化学系。
    9
    消化和肝病，Vall D'hebron 研究所，巴塞罗那自治大学，西班牙巴塞罗那。
    10
    巴塞罗那自治大学，西班牙贝拉特拉。

#
同等贡献。

    PMID：35377229 DOI：10.1128/spectrum.02149-21

抽象的

对接受抗病毒治疗的 HBV 感染患者 HBV DNA 和 RNA 水平的测量和解释支持了 HBV 疾病管理的目标。在这里，我们通过标准化和敏感的测定对来自幼稚和接受治疗的患者的随访样本中的循环 HBV RNA 进行量化，作为感染演变的标志物。 HBV DNA（用于 Cobas 6800/8800 自动罗氏分子系统的 HBV DNA）、RNA（用于 Cobas 6800/8800 的罗氏 HBV RNA 研究分析；罗氏）、HBeAg 和 HBsAg（Cobas 8000 的 Elycsys HBsAg 化学发光免疫分析；罗氏）和核心相关抗原（Lumipulse G 化学发光测定法；Fujirebio）水平在门诊医院环境中未经治疗或核苷（酸）治疗的 HBV 感染受试者队列中进行测量。未经治疗的人的 HBV DNA 水平比相应的 RNA 水平高 3.6 log10，并且在 5 年的观察中保持稳定。虽然在随访结束时，52 名接受治疗的患者中只有 5 名 DNA 水平低于定量下限 (10 IU/mL)，但 13 名 HBV RNA 水平持续高于该限值，其中 8 名 DNA 检测不到。在核心相关抗原检测不到的样本中，我们观察到 HBsAg 滴度中位数比 RNA 检测不到的样本高 2.7 倍（调整后的 P = 0.012）。可检测到的 HBV RNA 和不可检测的 HBV DNA 是 HBsAg 降至 ≤100 IU/mL 水平的负预测因子（P = 0.03）。在幼稚患者中，HBV DNA 和 RNA 之间的差异高于先前报道的。治疗期间HBV RNA迅速下降。然而，在某些情况下，即使经过多年的有效治疗后仍可检测到，这是 HBsAg 下降的负面预测因素。用于 Cobas 6800/8800 仪器的研究性 RNA 测定是一种敏感且标准化的方法，可用于 HBV 感染的一般管理。重要性 本研究的重点是通过使用标准化和灵敏的检测方法对循环 HBV RNA 进行定量。多亏了这个系统，我们观察到循环 HBV DNA 和 RNA 之间的差异比以前报道的要大。在接受治疗的患者中，HBV RNA 与 DNA 一起下降，尽管一些患者即使在多年成功的抗病毒治疗后仍表现出可检测的水平，表明存在持续的病毒转录。值得注意的是，当检测不到 HBV DNA 时，检测到病毒 RNA 是 HBsAg 降至 ≤100 IU/mL 水平的阴性预测因子。该检测方法对 HBV 患者管理以研究病毒转录和识别可能实现持续病毒抑制的治疗患者非常有帮助。

关键词：乙肝；乙肝病毒核糖核酸；逆转录聚合酶链反应；抗病毒治疗；生物监测。

StephenW

Standardized Hepatitis B Virus RNA Quantification in Untreated and Treated Chronic Patients: a Promising Marker of Infection Follow-Up
Maria Francesca Cortese  1   2   3 , Mar Riveiro-Barciela  3   4 , David Tabernero  2   3 , Francisco Rodriguez-Algarra  5 , Adriana Palom  4 , Sara Sopena  2 , Ariadna Rando-Segura  2   6 , Luisa Roade  4 , Alison Kuchta  7 , Roser Ferrer-Costa  8 , Josep Quer  3   9 , Beatriz Pacin  1   2   3 , Marta Vila  1   2 , Rosario Casillas  1   2 , Selene Garcia-Garcia  1   2   3 , Rafael Esteban  3   4 , Tomás Pumarola  6   10 , Maria Buti #  3   4 , Francisco Rodriguez-Frias #  1   2   3
Affiliations
Affiliations

    1
    Clinical Biochemestry, Vall D'hebron Research Institute, Universitat Autònoma de Barcelona, Barcelona, Spain.
    2
    Biochemistry and Microbiology, Liver Pathology Unit, Vall D'hebron University Hospital, Universitat Autònoma de Barcelona, Barcelona, Spain.
    3
    Centro De Investigación Biomédica En Red, Enfermedades Hepáticas y Digestvas (CIBERehd), Instituto De Salud Carlos III, Madrid, Spain.
    4
    Liver Unit, Internal Medicine Department, Vall D'hebron University Hospital, Universitat Autònoma de Barcelona, Barcelona, Spain.
    5
    The Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom.
    6
    Virology Unit, Microbiology Department, Vall D'hebron University Hospital, Universitat Autònoma de Barcelona, Barcelona, Spain.
    7
    Roche Molecular Systems, Inc., Pleasanton, California, USA.
    8
    Department of Biochemistry, Vall D'Hebron University Hospital, Barcelona, Spain.
    9
    Digestive and Liver Disease, Vall D'hebron Research Institute, Universitat Autònoma de Barcelona, Barcelona, Spain.
    10
    Universitat Autònoma de Barcelona, Bellaterra, Spain.

#
Contributed equally.

    PMID: 35377229 DOI: 10.1128/spectrum.02149-21

Abstract

The measurement and interpretation of HBV DNA and RNA levels in HBV infected patients treated with antiviral therapy supports the objective of HBV disease management. Here, we quantified circulating HBV RNA through a standardized and sensitive assay in follow-up samples from both naive and treated patients as a marker of infection evolution. HBV DNA (HBV DNA for use in Cobas 6800/8800 Automated Roche Molecular Systems), RNA (Roche HBV RNA Investigational Assay for use in the Cobas 6800/8800; Roche), HBeAg and HBsAg (Elycsys HBsAg chemiluminescence immunoassay by Cobas 8000; Roche), and core-related antigen (Lumipulse G chemiluminescence assay; Fujirebio) levels were measured in cohorts of untreated or nucleos(t)ide treated, HBV-infected subjects in an outpatient hospital setting. HBV DNA levels in untreated people were 3.6 log10 higher than corresponding RNA levels and were stable over 5 years of observation. While only five of 52 treated patients had DNA levels below the lower limit of quantification (10 IU/mL) at the end of follow-up, 13 had HBV RNA levels persistently above this limit, including eight with undetectable DNA. In samples with undetectable core-related antigen we observed a median HBsAg titer 2.7-fold higher than in samples with undetectable RNA (adjusted P = 0.012). Detectable HBV RNA with undetectable HBV DNA was a negative predictor of HBsAg decrease to a level ≤100 IU/mL (P = 0.03). In naive patients the difference between HBV DNA and RNA was higher than previously reported. HBV RNA rapidly decreased during treatment. However, in some cases, it was detectable even after years of effective therapy, being a negative predictor of HBsAg decrease. The investigational RNA assay for use on the Cobas 6800/8800 instruments is a sensitive and standardized method that could be applied in general management of HBV infection. IMPORTANCE This study focused on the quantification of circulating HBV RNA by using a standardized and sensitive assay. Thanks to this system we observed a higher difference between circulating HBV DNA and RNA than previously reported. In treated patients, HBV RNA decreased together with DNA, although some patients presented detectable levels even after years of successful antiviral treatment, suggesting a persistent viral transcription. Of note, the detection of viral RNA when HBV DNA is undetectable was a negative predictor of HBsAg decrease to a level ≤100 IU/mL. This assay could be extremely helpful in HBV patients management to study viral transcription and to identify those treated patients that may achieve sustained viral suppression.

Keywords: HBV; HBV-RNA; RT-PCR; antiviral treatment; biomonitoring.