Abstract：
OBJECTIVE: 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.
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.
摘要：
目的：慢性丁型肝炎（CHD）的管理需要可靠的HDVRNA定量测试。该研究的目的是比较两种提取方法，以定量未治疗和bulevirtide（BLV）治疗的CHD患者中的HDVRNA。
方法：在单中心研究中测试了未经治疗和BLV治疗的CHD患者的冷冻血清的HDVRNA水平（Robogene2.0，莱比锡,德国；LOD6IU/mL），具有两种提取方法：手动（INSTANT病毒RNA/DNA试剂盒；RoboscreenGmbH，莱比锡,德国)与自动(EZ1DSP病毒试剂盒；Qiagen，希尔登,德国)。BLV治疗的患者在基线和治疗期间取样。
结果：从157例冠心病（139例未经处理，对18例BLV治疗的)患者进行了分析：年龄51岁(28-78岁)，59％的男性，90%的欧洲血统，60%肝硬化,ALT85(17-889)U/L,HBsAg3.8（1.7-4.6）LogIU/mL，81%HBVDNA检测不到，98%HDV基因型1。手动与自动提取的HDVRNA中位数为4.53（.70-8.10）对3.77（.70-6.93）LogIU/mL（p<.0001）。手工提取在31（20％）患者中报告了相似的HDVRNA水平，119人中较高(76%)[60人中+5和+1log10；59人中>+1log10]，7人中较低(4%)。在18名接受BLV治疗的患者中，在第16周和第24周，两种检测方法的HDVRNA结论：Robogene2.0对HDVRNA的定量受提取方法的影响，手动提取是1日志更敏感。
方法：在单中心研究中测试了未经治疗和BLV治疗的CHD患者的冷冻血清的HDVRNA水平（Robogene2.0，莱比锡,德国；LOD6IU/mL），具有两种提取方法：手动（INSTANT病毒RNA/DNA试剂盒；RoboscreenGmbH，莱比锡,德国)与自动(EZ1DSP病毒试剂盒；Qiagen，希尔登,德国)。BLV治疗的患者在基线和治疗期间取样。
结果：从157例冠心病（139例未经处理，对18例BLV治疗的)患者进行了分析：年龄51岁(28-78岁)，59％的男性，90%的欧洲血统，60%肝硬化,ALT85(17-889)U/L,HBsAg3.8（1.7-4.6）LogIU/mL，81%HBVDNA检测不到，98%HDV基因型1。手动与自动提取的HDVRNA中位数为4.53（.70-8.10）对3.77（.70-6.93）LogIU/mL（p<.0001）。手工提取在31（20％）患者中报告了相似的HDVRNA水平，119人中较高(76%)[60人中+5和+1log10；59人中>+1log10]，7人中较低(4%)。在18名接受BLV治疗的患者中，在第16周和第24周，两种检测方法的HDVRNA
