某国产HIV-1病毒载量检测试剂的检测性能评估

目的对1型艾滋病病毒(HIV-1)的国产病毒载量检测试剂进行检测性能评估。方法应用系列稀释的定值质控品分析某国产病毒载量检测试剂的检测线性度和精密度,应用81份HIV-1感染阳性样本和30份阴性样本比较该国产病毒载量检测试剂与进口TaqMan病毒载量检测试剂检测结果的相关性和一致性。结果国产病毒载量检测试剂对系列浓度定值质控品检测结果的线性相关系数为R2=0.997 0。对不同浓度定值质控品检测结果的批间和批内检测变异系数均10%。与进口TaqMan病毒载量检测试剂相比较,两种检测试剂对临床样本检测结果之间的相关性系数为R2=0.792 3。比较两种试剂检测结果偏差的Bland-Altman分析显示偏差均值为-0.01 Log(拷贝/mL)。两种试剂判定阳性和阴性样本的检测符合率为100%。以1 000拷贝/mL为检测线时,两种试剂检测结果的符合率为93.83%;以5 000拷贝/mL为检测线时,两种试剂检测结果的符合率为91.36%。结论某国产病毒载量检测试剂具有较好的检测线性和精密度,与进口TaqMan病毒载量检测试剂相比具有较好的相关性和一致性,整体检测性能良好可适用于大部分临床检测需求。     

两种HIV-1 RNA定量检测方法的临床对比研究

目的比较达安基因艾滋病病毒1型(HIV-1)核酸检测Ⅱ代(简称达安基因)和罗氏Cobas TaqMan HIV-1Test Version 2.0(简称罗氏Cobas TaqMan)两种试剂,检测HIV-1血浆病毒载量的相关性及一致性。方法采用达安基因试剂和罗氏Cobas TaqMan试剂,平行检测663份HIV-1感染者血浆病毒载量,采用配对t检验、相关分析和Bland-Altman等方法进行统计分析。结果罗氏Cobas TaqMan与达安基因试剂检测结果高于检测下限的比例一致(分别为65.31%和65.76%)。对于病毒载量低于1 000IU/mL的样本,二者的差异无统计学意义(P=0.856 8)。罗氏Cobas TaqMan与达安基因试剂检测结果具有很强的相关性(r=0.962 0,P0.000 1)。此外,Bland-Altman分析两种方法检测病毒载量的结果具有较高的一致性。结论达安基因和罗氏Cobas TaqMan试剂,检测HIV-1血浆病毒载量的结果具有较好的相关性和一致性。     

病毒载量试验对“HIV-1抗体不确定”病例诊断的效果评价北大核心CSCD

目的评价病毒载量（VL）试验对＂艾滋病病毒Ⅰ型（HIV-1）抗体不确定＂病例诊断的价值。方法对一组106例＂HIV-1抗体不确定＂病例进行定期随访,检测其VL与抗体,比较两种检测策略对这类病例的诊断效果。结果共106例HIV-1抗体不确定病例,均在一周内进行了COBAS TaqMan HIV-1Test v2.0病毒载量检测,56例≥100 000拷贝/mL,31例在5 000~100 000拷贝/mL之间,6例在20~5 000拷贝/mL之间,13例核酸未检出。常规随访抗体检测中,27例（25.5%）在4周内明确诊断,59例（55.7%）超过4周复查明确诊断,12例（11.3%）失访,5例（4.7%）死亡,3例多次随访结果仍为＂HIV-1抗体不确定＂。结论 VL试验能快速准确鉴别不确定结果,缩短随访复查的时间,是对现有抗体补充试验复查检测策略的重要补充和完善。     

一种国产HIV核酸定量试剂与进口试剂的对比研究

目的对比研究国产1型艾滋病病毒(HIV-1)核酸定量试剂与两个进口试剂的相关性和差异,以期应用国产核酸定量试剂评价中国抗病毒治疗患者病毒学效果的可行性。方法应用美国国家卫生研究院病毒学质量评价项目(VQA)发放的HIV-1核糖核酸(RNA)血清样品,比较一种国产核酸定量试剂与国内市场常规使用的两个进口试剂,在测定病毒载量上的相关性和一致性。对国产和进口核酸定量试剂检测HIV-1病毒载量的结果进行配对t检验,线性回归分析和Bland-Altman分析。结果国产试剂和两个进口试剂的检测结果的线性相关系数R2分别为0.94、0.93(P0.000 1);Bland-Altman分析三种试剂病毒载量的结果具有较高的一致性,国产试剂定量值与VQA病毒载量值线性相关系数R2=0.97,P0.000 1,定量单位转换系数为1IU/mL=0.79拷贝/mL。结论国产HIV-1核酸定量试剂与进口试剂在定量检测结果上有较好的相关性和一致性,而且国产试剂在价格上具有优势,在一定范围内可以应用于抗病毒治疗效果的评价。     

bDNA和NASBA法定量检测HIV-1病毒载量的比较研究

目的比较分支DNA杂交实验(branched DNA,bDNA)和核酸序列扩增实验(Nucleic acid sequence-based am-plification,NASBA)两种方法检测人类免疫缺陷病毒1型病毒(HIV-1)病毒载量间的一致性。方法对25例HIV感染者/艾滋病(Acquired immune deficiency syndrome,AIDS)患者血浆标本同时用bDNA法和NASBA法检测HIV-1病毒载量,并用流式细胞术检测患者外周血CD4+、CD8+T淋巴细胞。结果bDNA法及NASBA法测得HIV-1病毒载量平均值分别为(4.398±0.580)log拷贝数/ml和(4.488±0.602)log拷贝数/ml,差异无统计学意义(t=1.210,P>0.05);两种方法检测出的病毒载量呈显著直线相关(r=0.8004,P<0.001),且与患者的CD4+T细胞数及CD4+/CD8+比值均呈显著直线负相关。结论bDNA法和NASBA法检测HIV-1病毒载量具有高度一致性,在实际工作中均可选用。     

Nuclisens EasyQ HIV-1和COBAS AMPLICOR HIV-1 MONITORTM Test检测集合在HIV-1“窗口期”诊断的比较应用

目的将Nuclisens EasyQ HIV-1方法与COBAS AMPLICOR HIV-1MONITORTM Test方法检测集合的结果进行比较,并将集合HIV RNA Nuclisens EasyQ方法应用于艾滋病病毒Ⅰ型(HIV-1)"窗口期"的检测。方法采用50:1、10:1二级集合HIV RNA COBAS和EasyQ方法进行检测。结果 (1)两种方法在54份定值血清和质控血浆的阳性检出率均为100%,方法的一致性为94.44%,两方法之间具有显著相关性,相关系数为r=0.855。(2)检测感染者配偶、静脉吸毒者(IDU)及男男性行为人群(MSM)样本集合2 426人份,两种方法检测结果一致,发现3例"窗口期"感染者。结论集合HIV RNA Nuclisens EasyQ方法具有很好的实验性能,可应用于HIV高危人群的"窗口期"检测。     

保存时间对血浆样本HIV-1病毒载量测定值的影响研究

目的观察保存时间对血浆样本艾滋病病毒Ⅰ型（HIV-1）病毒载量测定值的影响。方法应用HIV-1核酸序列扩增系统技术（NASBA）对87份-20℃保存的HIV-1抗体阳性血浆样本分组，分别在第0、3、6、11、12、13、14个月进行HIV-1病毒载量检测。结果-20℃保存3个月的样本病毒载量无明显降低，平均下降0．210Log值（P〉0．05），6个月以后的样本病毒载量明显降低，平均下降0．256Log值（P〈0．05），11、12、13个月以后的样本病毒载量明显降低，平均下降大于0．428Log值（P〈0．05），但病毒载量下降幅度与原始病毒载量无关（P〉0．05）。结论血浆样本在-20℃条件下保存6个月以上HIV-1RNA病毒载量水平降低明显，已不能真实反映原始样本病毒载量水平，用于病毒载量检测的血浆样本在-20℃冰柜中不宜超过3个月。     

云南省2008-2012年HIV-1耐药基因型检测情况

目的了解云南省2008—2012年艾滋病病毒Ⅰ型（HIV-1）耐药基因检测及临床应用情况。方法采用反转录套式聚合酶链反应（RT-nested—PCR）方法,检测病毒抑制失败病人血浆HIV-1耐药基因型。结果截止2012年12月31日,云南省累计治疗艾滋病（AIDS）病人38055人。2008—2012年,HIV1病毒载量（VL）检测率逐年提高（54．0％～95．2％）。HIV-1VL〈50拷贝／mL者占88．7％,VL在51～1000拷贝／mL者占3．4％,VL〉1000拷贝／mL者占7．9％。对VL〉1000拷贝／mL病人血浆进行HIV-1耐药基因突变检测,共检测3766人次,获得可用序列2586例。其中未检出耐药突变的占46．2％（1196／2586）,发生耐药突变的占53．8％（1390／2586）。以2012年末累计治疗人数为基数,计算得到抗病毒治疗（HAART）人群的耐药发生率为3．7％（1390／38055）。16个地市中除丽江和迪庆两地未发现耐药毒株外,其余各地均有不同程度的耐药发生。核苷类反转录酶抑制剂（NRTI）、非核苷类反转录酶抑制剂（NNRTI）和蛋白酶抑制剂（PI）的突变率分别为63．3％、95．1％和8．5％。NRTI突变位点的前3位依次为M184V／I（52．7％）、T215Y／F（11．7％）、D67N（10．2％,）,NNRTI前3位是K103N／S（37．8％）,G190A／S／E（24．4％）,Y181C／I／V（22．0%）,PI出现最多的是L33F,其次还有M46I、V82A。结论云南省抗病毒治疗病毒学失败病人中,以NNRTI耐药最为多见,其次为NRTI,PI较少,各地、市HIV耐药发生存在一定差异。HIV耐药基因检测有助于科学抉择治疗方案,在提高HAART的效果的同时,有效地节约匮乏的药物资源,减少耐药毒株的流行传播。     

国产实时荧光定量核酸检测试剂测定HIV病毒载量

目的在目前进口试剂较昂贵的情况下,探讨国产实时荧光定量核酸检测试剂用于检测艾滋病病毒(HIV)病毒载量的可行性。方法2004年从全国4个省的HIV感染者/艾滋病(AIDS)患者采集110份样本,每份样本均同时使用生物梅里埃公司NASBA与深圳匹基生物公司实时荧光定量PCR两种方法测定血浆中HIV RNA含量,比较两种方法所得数据间的关系。结果HIV样本病毒载量处于3.5×103-1.0×105拷贝/ml范围时,一致性较好,存在一定线性关系;在小于103拷贝/ml范围内时,两种方法检测结果基本一致;当病毒载量处于1.0×103-3.5×103拷贝/ml范围时,两种方法检测结果偏差较大;病毒载量处于大于105拷贝/ml的范围时,虽然两种检测方法数值之间无相关性,但均显示处于105以上的较高数值范围。结论NASBA与实时荧光定量PCR两种检测方法在3.5×103-1.0×105拷贝/ml范围内有着高度的相关性,国产实时荧光定量核酸检测试剂检测结果与国际上较常用的病毒载量检测方法的结果之间的差异已经缩小,在低载量的检测敏感性还需要进一步优化。总体来说,使用国产实时荧光定量核酸检测试剂可以初略定量血浆中HIV RNA含量。     

血浆HIV-1 RNA与干血斑HIV-1 DNA基因型耐药检测比较

目的 通过对不同病毒载量的同一研究对象同时进行血浆HIV-1 RNA与干血斑HIV-1 DNA的基因型耐药检测并对结果进行比较,探讨HIV-1 DNA用于耐药检测的可行性和用途。方法 2021年12月采集云南省、广西壮族自治区及新疆维吾尔自治区接受ART后1年以上的HIV/AIDS患者静脉血5 mL,EDTA抗凝,分离血浆和制备成干血斑样本,分别提取病毒DNA和RNA进行pol区扩增,比较两种方法的扩增效率;并对同时扩增成功的序列利用MEGA7构建系统进化树并分析序列一致性。利用斯坦福大学HIV耐药数据库进行耐药位点分析,比较耐药性结果。结果 209例样本中,来自云南82份（39.2%）,来自广西壮族自治区69份（33.0%）,来自新疆维吾尔自治区58份（27.8%）。105例VL<20拷贝/mL,25例20拷贝/mL≤VL<200拷贝/mL,42例200拷贝/mL≤VL<1 000拷贝/mL,37例VL≥1 000拷贝/mL。不同病毒载量的样本分类中,使用血浆中HIV-RNA pol区扩增成功率分别为12.4%、28.0%、69.0%、89.2%;使用干血斑中HIV...     

Evaluation of the COBAS ampliprep/COBAS TaqMan system for quantification of human immunodeficiency virus type 1 (HIV-1) RNA by real-time PCR

BACKGROUND: Viral load quantification is standard for monitoring HIV-1 therapy and is crucial in deciding whether to switch or to continue a current antiretroviral regimen. In Japan, serum is widely adapted as a specimen of the HIV-1 viral load quantification assay. METHODS: We evaluated an emerging HIV-1 RNA quantification of the COBAS AmpliPrep/COBAS TaqMan HIV-1 Test (COBAS TaqMan). The test was evaluated for matrix equivalence between plasma and serum and for correlation with the AMPLICOR HIV-1 Monitor Test v1.5 (Amplicor) for HIV-1 RNA quantification. RESULTS: The test result from serum specimens showed good correlation with test results from plasma specimens. HIV-1 RNA quantification results using serum specimens correlated well with those obtained by both ultrasensitivity assay and standard Amplicor assay. CONCLUSIONS: The fully automated COBAS AmpliPrep/COBAS TaqMan HIV-1 Test meets requirements for a wide dynamic range and reliable quantification of HIV-1 RNA in serum clinical specimens.     

Comparative evaluation of the COBAS AmpliPrep/COBAS TaqMan HIV type 1 test (CAP/CTM) and VERSANT HIV type 1 RNA 3.0 assay (bDNA) for quantifying HIV type 1 viral loads in China

In the present study, 277 clinical samples from untreated and treated HIV-1-infected patients with different clades were used to assess the agreement between the COBAS AmpliPrep/COBAS TaqMan HIV-1 test (CAP/CTM) and VERSANT HIV-1 RNA 3.0 Assay (bDNA). A qualitative comparison of the results of the two assays showed concordance for 255 positive and 15 negative samples (94.95%, kappa = 0.798). However, seven samples with viral loads close to the lower limit of detection for CAP/CTM were negative by bDNA. A significant correlation (r = 0.881, p < 0.001) was observed for 253 samples with viral loads within the dynamic ranges of the two assays, and Bland-Altman analysis showed good agreement (96.05%) between the two assays for these 253 samples [mean (+/-2 SD), 0.389(-0.385, 1.163)]. Furthermore, ART drugs had no impact on the performances of the two assays. For samples with different clades predominant in China, the fitted regression line differed significantly from the line of equality, although significant correlations (r = 0.850-0.891, p < 0.001) and good agreements (92.86-97.25%) were found for the two assays. The mean differences for clade B' and BC samples were significant (p < 0.01). Good precision for clade B' samples was achieved for the CAP/CTM (CV: 20.73%) and bDNA (CV: 12.19%) assays. Furthermore, for clades B', BC, and AE, both assays exhibited good linearities (r = 0.9773-0.9998). Thus, the CAP/CTM and bDNA assays could be useful for quantifying HIV-1 RNA in routine clinical samples and monitoring viral loads in treated and untreated HIV-infected patients in China.     

Evaluation of a new real-time PCR assay kit for quantification of human immunodeficiency virus RNA in plasma

We evaluated the use of COBAS TaqMan HIV-1 for highly pure system (COBAS TaqMan) recently developed as a second-generation quantification of human immunodeficiency virus type 1 (HIV-1) RNA. The linearity, sensitivity, reproducibility, and effects of possibly interfering materials were examined and results compared to those of AMPLICOR HIV-1 MONITOR Test version 1.5 (AMPLICOR) using 6 virus isolates that were all different subtypes. Excellent linearity was obtained at 1.67 x 10(2)-1.73 x 10(6) copies/mL (r2 = 0.991). Sensitivity was 40 copies/mL at a 100% hit rate. Intraexperimental CVs were 27.4-50.8%, and interexperimental CVs were 29.3-81.5%. Although COBAS TaqMan showed an excellent titer correlation with AMPLICOR (r2 = 0.960), mean titer obtained with COBAS TaqMan was 3.1 times higher than that with AMPLICOR (p = 0.002), and 7.1 times higher in a sample of subtype C, suggesting discrepancies in HIV-1 RNA quantification between the two kits. This point should be noted when AMPLICOR is replaced by COBAS TaqMan.     

2种PCR试剂盒定量检测血清HBV DNA含量的比较

目的探讨AMPLIPREP-COBAS TAQMAN法(罗氏COBAS法)和北京鑫诺美迪PCR-荧光探针"一管法"检测血清HBV DNA含量的性能差异。方法采用2种方法同步检测175例乙型肝炎患者血清样本,并将黄疸、溶血和脂血标本作为干扰样本,分析2种方法的相关性、一致性和抗干扰性。取一已知定量为2.24×109IU/ml的标本,用阴性血清做1+9(即1∶10)的稀释,并依次稀释至2.24×10 IU/ml,比较2种方法的线性范围和灵敏性的差异。结果 175份均有数值的标本中,2种试剂检测结果比较差异无统计学意义。2种试剂对黄疸、溶血和脂血标本的定量值影响不大。对于HBV DNA1.70×108IU/ml的样本,罗氏COBAS法结果只显示1.70×108IU/ml,而"一管法"无须稀释仍能准确定量;对于HBV DNA1.00×102IU/ml的样本,"一管法"仅能检测到病毒,而罗氏COBAS法的稳定性和线性更好。结论 PCR-荧光探针"一管法"与进口罗氏COBAS法具有良好的一致性,且省时、省力,价格低廉,适合在我国推广应用。     

两种实时荧光定量聚合酶链反应检测血清HCV RNA的比较

目的 比较全自动病毒载量检测系统(COBAS TaqMan)和国产荧光定量PCR试剂盒对血清HCV RNA载量的检测结果,探讨两种检测方法在临床诊断和治疗中的应用价值.方法 收集26例慢性丙型肝炎患者抗病毒治疗前和治疗过程中2、4、8、12、24、36和48周的系列血标本,共168份,采用COBAS TaqMan 48全自动分析系统和广州某国产TaqMan实时PCR试剂盒分别检测系列血清中的HCV RNA载量.统计学处理采用x2检验和t检验.结果 当血清HCV RNA≥1×104IU/mL时(0周),COBAS检测和国产试剂盒均能很好测定HCV载量,而且国产试剂盒检测值为1.35×107IU/mL高于COBAS检测值2.21×106IU/mL,差异有统计学意义(t=2.05,P<0.05);血清HCV RNA<1×104IU/mL时(2～48周),COBAS检测出的HCV阳性率为21.4%(30/140),远高于国产试剂盒的1.4%(2/140),差异有统计学意义(t=3.66,P<0.01);治疗4周时,COBAS检测26例患者中14例血清HCV为阳性,12例病毒载量低于检测下限,获得快速病毒学应答(RVR);国产试剂盒检测结果为3例血清HCV为阳性,23例获得RVR.COBAS与国产试剂盒梧比,转阴率差异有统计学意义(x2=10.575,P<0.01).治疗12周时,COBAS检测完全早期病毒学应答(cEVR)率为95.7%(22/23),国产试剂盒检测的cEVR为100%(17/17),差异无统计学意义(x2=0.726,P>0.05).结论 国产荧光定量PCR试剂盒可用于HCV疑似患者的筛查和高HCVRNA载量者的确诊,对于低HCV RNA载量的疑似患者和抗病毒治疗过程中HCV载量的检测,COBAS则更为敏感.

Abstract：

Objective To compare the plasma hepatitis C virus(HCV)RNA levels detected by the fully automated viral load detection system(COBAS TaqMan)and the national real-time quantitative polymerase chain reaction(PCR)kit,and to investigate the clinical application value of these two methods in clinical practice.Methods A total of 168 serial plasma samples collected from 26 patients with chronic hepatitis C(CHC)before and at week 2,4,12,24,36 and 48 of antiviral treatment were detected by both COBAS Taqman 48 analyzing system and the national real-time quantitative PCR kit.The results of two methods were compared by chi square test and t test.Resnlts Both COBAS and national kit showed great positive detecting results when HCV RNA≥1×104IU/mL(at week O),and the virus load value detected by national kit was significantly higher than that detected by COBAS(t=2.05,P<0.05).However,when HCV RNA<1×104(at week 2-48),the positive rate of HCV detected by COBAS was significantly higher than that detected by national kit (t=3.66,P<0.01).At week 4 of treatment,the rapid virological response(RVR)rate was 46.2 % (12/26)detected by COBAS,while that was 88.5%(23/26)detected by national kit,and the difference was significant(x2=10.575,P<0.01).At week 12 of treatment,the complete early virological response(cEVR)was 95.7%(22/23)detected by COBAS,while that was 100%(17/17)detected by national kit,and the difference was not significant(x2=0.726,P>0.05).Conclusions The national TaqMan real-time quantitative PCR kits could be used to screen the suspected cases of HCV infecrion and to diagnose CHC cases with high HCV virus load.COBAS detection is more sensitive in cases with low HCV virus load and in on-treatment monitor during anti-HCV therapy.     

Evaluation of COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen HBV DNA, HCV RNA and HIV-1 RNA amplification and detection

BACKGROUND AND OBJECTIVES: This report describes the evaluation of the COBAS AmpliPrep instrument for fully automated generic nucleic acid extraction in conjunction with hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA, and human immunodeficiency virus (HIV)-1 RNA COBAS AmpliScreen amplification and detection using serial dilutions of the WHO international standards (IS) and the PeliCheck reference panels. MATERIALS AND METHODS: Serial diluted samples of the WHO IS and the PeliCheck reference panels were tested 24 times to determine the HBV DNA, HCV RNA and HIV-1 RNA detection limits by Probit analysis. The existence and extent of cross-contamination were assessed by testing alternating high titre HBV DNA-positive and -negative samples. The specificity of the AmpliPrep-AmpliScreen test for HBV was determined by testing 232 minipools consisting of six donations, all negative for HCV/HIV-1 nucleic acid testing (NAT) and HBsAg. In addition, a HBV genotypes A-G panel was tested. RESULTS: The respective 95% detection limits (and 95% CI) on the WHO IS and on the PeliCheck reference panels were 6.7 (4.3-13) IU/ml and 123 (68-301) gEq/ml for HBV DNA, 23 (11-106) IU/ml and 126 (84-233) gEq/ml for HCV RNA, and 187 (108-422) IU/ml and 183 (108-434) gEq/ml for HIV-1 RNA. Based on the WHO IS and the PeliCheck reference panels, no significant differences in sensitivity for HBV and HCV were found between AmpliPrep and the licensed MultiPrep extraction method. The sensitivity of AmpliPrep-AmpliScreen for HIV-1 was probably twofold lower as compared to the MultiPrep-AmpliScreen method. No cross contamination was observed. All 232 minipools were HBV NAT-negative. The AmpliPrep-AmpliScreen test for HBV detected HBV genotypes A-G with equal sensitivity. CONCLUSIONS: The AmpliPrep instrument combined with the AmpliScreen assays for HBV, HCV and HIV-1 is robust and suitable for NAT donor screening. The sensitivity criteria for HIV-1 and HCV as defined by the Paul Ehrlich Institute and the Food and Drug Administration for minipool NAT screening are met by this system. SINGLE SENTENCE SUMMARY: Generic COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen detection for HBV, HCV and HIV-1.     

Implications of HCV RNA level at week 4 of direct antiviral treatments for hepatitis C

We aimed to determine whether the HCV viral load after four weeks of treatment (W4VL) with direct‐acting antiviral agents (DAAs) predicts sustained virologic response (SVR) in a real‐world clinical setting. We identified 21 095 patients who initiated DAA‐based antiviral treatment in the national Veterans Affairs (VA) healthcare system from 01/01/2014 to 06/30/2015. Week 4 viral load was categorized as undetectable, detectable below quantification (DBQ), detectable above quantification (DAQ) with viral load ≤42 IU/mL and DAQ with viral load >42 IU/mL. Week 4 viral load was undetectable in 36.1%, detectable below quantification in 45.6%, DAQ ≤42 in 9.3%, DAQ >42 in 9.1%. Detectable above quantification was much more common and undetectable week 4 viral load much less common when tested with the Abbott RealTime HCV assay vs the Roche COBAS AmpliPrep/COBAS TaqMan Version 2 assay. Compared to patients with undetectable week 4 viral load (SVR=93.5%), those with detectable below quantification (SVR=91.8%, adjusted odds ratio [AOR] 0.79, P‐value=.001), DAQ ≤42 (SVR=90.0%, AOR 0.63, P‐value<.001) and DAQ >42 (SVR=86.2%, AOR 0.52, P‐value<.001) had progressively lower likelihood of achieving SVR after adjusting for baseline characteristics and treatment duration. Among genotype 1‐infected patients who were potentially eligible for 8‐week sofosbuvir/ledipasvir monotherapy, we did not find evidence that treatment for 12 weeks instead of 8 weeks was associated with higher SVR, even among those with detectable above quantification. In summary, DBQ and DAQ W4VL are very common in real‐world practice, contrary to what was reported in clinical trials, and strongly predict reduced SVR across genotypes and clinically relevant patient subgroups. Whether and how week 4 viral load results should influence treatment decisions requires further study.