罗氏 2.0 系统检测 HDV 患者血清 HDV RNA 的结果受提取方法影响显著。
Quantification of serum HDV RNA by Robogene 2.0 in HDV patients is significantly influenced by the extraction methods.
机构信息
Division of Gastroenterology and Hepatology, Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy.
Virology Unit, Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy.
出版信息
Liver Int. 2024 Mar;44(3):831-837. doi: 10.1111/liv.15795. Epub 2024 Jan 22.
BACKGROUND AND AIM
Management of chronic hepatitis delta (CHD) requires reliable tests for HDV RNA quantification. The aim of the study was to compare two extraction methods for the quantification of HDV RNA in untreated and bulevirtide (BLV)-treated CHD patients.
METHODS
Frozen sera from untreated and BLV-treated CHD patients were tested in a single-centre study for HDV RNA levels (Robogene 2.0, Roboscreen GmbH, Leipzig, Germany; LOD 6 IU/mL) with two extraction methods: manual (INSTANT Virus RNA/DNA kit; Roboscreen GmbH, Leipzig, Germany) versus automated (EZ1 DSP Virus Kit; Qiagen, Hilden, Germany). BLV-treated patients were sampled at baseline and during therapy.
RESULTS
Two hundred sixty-four sera collected from 157 CHD (139 untreated, 18 BLV-treated) patients were analysed: age 51 (28-78), 59% males, 90% of European origin, 60% cirrhotics, ALT 85 (17-889) U/L, HBsAg 3.8 (1.7-4.6) Log IU/mL, 81% HBV DNA undetectable, 98% HDV genotype 1. Median HDV RNA was 4.53 (.70-8.10) versus 3.77 (.70-6.93) Log IU/mL by manual versus automated extraction (p < .0001). Manual extraction reported similar HDV RNA levels in 31 (20%) patients, higher in 119 (76%) [+.5 and +1 log10 in 60; > +1 log10 in 59] and lower in 7 (4%). Among 18 BLV-treated patients, rates of HDV RNA < LOD significantly differed between the two assays at Weeks 16 and 24 (0% vs. 22%, p = .02; 11% vs. 44%, p = .03), but not at later timepoints. By contrast, virological response rates were similar.
CONCLUSIONS
Quantification of HDV RNA by Robogene 2.0 is influenced by the extraction method, the manual extraction being 1 Log more sensitive.
背景与目的
慢性丁型肝炎（CHD）的管理需要可靠的 HDV RNA 定量检测方法。本研究旨在比较两种提取方法在未经治疗和丁型肝炎病毒（HBV）衣壳抑制剂（BLV）治疗的 CHD 患者中检测 HDV RNA 的效果。
方法
在一项单中心研究中，使用两种提取方法（手动 INSTANT Virus RNA/DNA 试剂盒；Roboscreen GmbH，莱比锡，德国；LOD 6 IU/mL）和 Robogene 2.0（Roboscreen GmbH，莱比锡，德国；LOD 6 IU/mL）检测了来自 157 例 CHD（139 例未经治疗，18 例 BLV 治疗）患者的 264 份冷冻血清中的 HDV RNA 水平。BLV 治疗的患者在基线和治疗期间进行了采样。
结果
分析了来自 157 例 CHD（139 例未经治疗，18 例 BLV 治疗）患者的 264 份血清：年龄 51（28-78）岁，男性占 59%，90%为欧洲人，60%为肝硬化患者，丙氨酸氨基转移酶（ALT）85（17-889）U/L，HBsAg 3.8（1.7-4.6）Log IU/mL，81%HBV DNA 不可检测，98%HDV 基因型为 1 型。手动提取与自动提取相比，HDV RNA 中位数分别为 4.53（.70-8.10）和 3.77（.70-6.93）Log IU/mL（p<.0001）。手动提取报告在 31 例（20%）患者中 HDV RNA 水平相似，在 119 例（76%）患者中更高[+0.5 和+1 log10 有 60 例；+1 log10 有 59 例]，在 7 例（4%）患者中更低。在 18 例 BLV 治疗的患者中，两种检测方法在第 16 周和第 24 周时 HDV RNA<.LOD 的比例显著不同（0% vs. 22%，p=.02；11% vs. 44%，p=.03），但在后续时间点则没有差异。相比之下，病毒学应答率相似。
结论
Robogene 2.0 定量检测 HDV RNA 的结果受到提取方法的影响，手动提取比自动提取敏感 1 个对数。
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