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中华诊断学电子杂志 ›› 2013, Vol. 01 ›› Issue (01) : 10 -18. doi: 10.3877/cma.j.issn.2095-655X.2013.01.003

所属专题： 专题评论； 文献；

实验研究

新型丙型肝炎病毒抗体检测试剂Elecsys anti-HCV Ⅱ的性能评价与比较
杨瑞锋1, 管文莉1, 王茜1, 刘艳1, 魏来1,†()   
  1. 1. 100044　北京大学人民医院　北京大学肝病研究所，丙型肝炎和肝病免疫治疗北京市重点实验室
  • 收稿日期:2013-10-06 出版日期:2013-11-26
  • 通信作者: 魏来
  • 基金资助:
    国家"十二五"科技重大专项资助项目(2012ZX10002003); 北京大学人民医院研究与发展基金(RDC2012-06)

A newly developed hepatitis C virus screening assay, Elecsys anti-HCV Ⅱ assay: performance evaluation and comparison with other widely used assays

Ruifeng Yang1, Wenli Guan1, Qian Wang1, Yan Liu1, Lai Wei1,†()   

  1. 1. Peking University People′s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing 100044, China
  • Received:2013-10-06 Published:2013-11-26
  • Corresponding author: Lai Wei
  • About author:
    Corresponding author: Wei Lai, Email:
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引用本文：

杨瑞锋, 管文莉, 王茜, 刘艳, 魏来. 新型丙型肝炎病毒抗体检测试剂Elecsys anti-HCV Ⅱ的性能评价与比较[J]. 中华诊断学电子杂志, 2013, 01(01): 10-18.

Ruifeng Yang, Wenli Guan, Qian Wang, Yan Liu, Lai Wei. A newly developed hepatitis C virus screening assay, Elecsys anti-HCV Ⅱ assay: performance evaluation and comparison with other widely used assays[J]. Chinese Journal of Diagnostics(Electronic Edition), 2013, 01(01): 10-18.

目的

评价新上市的丙型肝炎病毒抗体(抗-HCV)筛查试剂Elecsys anti-HCV Ⅱ的性能，并与其他两种临床上广泛应用的同类试剂进行性能比较。

方法

使用罗氏Elecsys anti-HCV Ⅱ、雅培Architect anti-HCV和强生Vitros anti-HCV 3种试剂，平行检测4个HCV血清转换盘、861份临床常规样本、100份抗-HCV检测临界阳性样本及178份HIV感染患者样本。抗-HCV确诊试验为重组免疫印迹法(RIBA 3.0)或HCV RNA定量检测。此外，使用Elecsys anti-HCV Ⅱ检测203份不同HCV基因型样本以评价其基因型覆盖度。

结果

相比Architect和Vitros anti-HCV，Elecsys anti-HCV II可提前7～14d检测到HCV感染后抗-HCV的产生；在检测临床常规样本(包括抗-HCV临界阳性样本)时，Elecsys anti-HCV Ⅱ有100%的敏感度及良好的特异度；70.22%(125/178)HIV感染患者样本为HCV RNA阳性，Elecsys anti-HCV Ⅱ可检测出其中97.60%(122/125)样本中的抗-HCV；Elecsys anti-HCV Ⅱ可能低估3b型样本的抗-HCV水平。

结论

Elecsys anti-HCV Ⅱ可进一步缩短HCV感染后的检测窗口期，可灵敏、特异地检测临床样本包括免疫缺陷患者样本中的抗-HCV，适用于临床HCV感染的筛查，但对其检测特定HCV基因型样本的性能尚需纳入更多样本深入研究。

关键词: 肝炎病毒，丙型, 肝炎抗体，丙型, 基因型
Background

The persistence of hepatitis C virus (HCV) infection remains as a major cause of liver cirrhosis and hepatocellular carcinoma. HCV infection is often asymptomatic, thus diagnosis relies heavily on clinical laboratory assays. As an "indirect" test, the anti-HCV screening assay fails to detect HCV infection in the seroconversion window and might report false-negative results in severely immunosuppressed populations.However, it is cheaper, simpler, and can provide results more rapidly. Therefore it remains the preferred method for screening HCV infection in most clinical laboratories.As a newly developed anti-HCV screening assay, the Elecsys anti-HCV Ⅱ assay has been recently launched. Objective To evaluate Elesys anti-HCV Ⅱ assay performance in Chinese populations and compare it with other widely used anti-HCV screening methods.

Methods

Four HCV seroconversion panels, 861 fresh consecutive serum samples under routine clinical conditions, 100 preselected serum samples with low positive anti-HCV results (tested before using the Vitros anti-HCV assay) and 178 samples from patients with HIV infection were tested the Elecsys anti-HCV Ⅱ assay and two comparator assays, Architect anti-HCV and Vitros anti-HCV assay. Confirmatory test was performed using recombinant immunoblot assay (RIBA) 3.0 or HCV RNA tests. Moreover, 203 samples with different HCV genotypes were assessed using the Elecsys anti-HCV Ⅱ assay to test its genotype inclusivity.

Results

The Elecsys anti-HCV Ⅱ assay detected HCV seroconversion 7-14 days earlier than the Architect anti-HCV and Vitros anti-HCV assay. It had 100% sensitivity and superior specificity in screening clinical routine samples. For the routine samples with positive anti-HCV by all the three screening assays, the S/Co values obtained using Elecsys anti-HCV Ⅱ did not correlate with those obtained Architect anti-HCV (P=0.13) Vitros anti-HCV (P=0.22) the S/Co values obtained correlated very well with a Pearson correlation coefficient of 0.97(P<0.01). Totally 73 preselected samples with low positive anti-HCV results remained anti-HCV positive when retested the Vitros anti-HCV assay, among which only 18 samples tested positive using Elecsys anti-HCV Ⅱ and 36 samples tested positive using the Architect anti-HCV assay. The Elecsys anti-HCV Ⅱ assay significantly distinguished the 73 anti-HCV borderline results; 55 negative samples tested using Elecsys anti-HCV Ⅱ had S/Co ratios 0.038-0.13, while 18 positive samples had S/Co ratios 13.58-135.6, respectively.Only one sample with an S/Co ratio of 13.58 by Elecsys anti-HCV Ⅱ was confirmed as RIBA indeterminate, while the other 17 samples with S/Co ≥20.0 were all RIBA positive. There were 161 out of 178 samples from HIV infected patients tested anti-HCV positive while 14 samples tested negative using all the three screening.The remaining three samples generated discordant results among the anti-HCV screening assays. The Elecsys anti-HCV Ⅱ assay, the Architect anti-HCV and the Vitros anti-HCV assay detected 97.6% (122/125) anti-HCV-positive samples from HIV-infected patients with HCV viremia. Using the Elecsys anti-HCV Ⅱ assay, however, the anti-HCV levels in the genotype 3b samples were slightly underestimated.

Conclusions

The Elecsys anti-HCV Ⅱ can further shorten the HCV seroconversion window due to its superior sensitivity. It is sensitive and specific enough for screening HCV infection in clinical routine samples as well.The Elecsys anti-HCV Ⅱ assay can be used together with either the Architect anti-HCV or the Vitros anti-HCV assay to improve the specificity in detecting samples with borderline positive anti-HCV S/Co ratios; this strategy will avoid unnecessary medical visits and psychological harm patients with a borderline positive anti-HCV result. The Elecsys anti-HCV Ⅱ assay is also suitable for testing samples from the immunocompromised patients, due to satisfactory sensitivity. Nevertheless, further investigation of its subtype inclusivity in a larger sample number might be warranted.

Key words: Hepatitis C virus, Hepatitis C Anti bodies, Genotype
图1 HCV感染临床常规样本及预选的抗体临界阳性样本的实验室检测流程图
表1 使用不同的抗-HCV筛查试剂检测4个HCV血清转换盘的结果
血清转换盘 初次采血后天数 HCV基因型 ALT(U/L) RIBA HCV RNA(拷贝/mL) Elecsys anti-HCV Ⅱ Architect anti-HCV Vitros anti-HCV
PHV 912-01 0 2b/3 13 NEG 4×104 8.51(POS) 0.30(NEG) 0.26(NEG)
PHV 912-02 4 2b/3 61 NEG >5×105 7.96(POS) 0.27(NEG) 0.25(NEG)
PHV 912-03 7 2b/3 298 IND 4×104 64.70(POS) 10.95(POS) 30.20(POS)
HCV 6212-1 0 1 55 NEG 7.50×105 2.96(POS) 0.04(NEG) 0.02(NEG)
HCV 6212-2 12 1 44 NEG 1.80×105 152.10(POS) 0.87(NEG) 0.73(NEG)
HCV 6212-3 14 1 41 NEG 8.50×104 163.80(POS) 1.98(POS) 1.30(POS)
HCV 6212-5 26 1 39 IND 4.90×104 107.30(POS) 8.14(POS) 5.10(POS)
HCV 6212-6 32 1 23 IND 8.20×104 105.40(POS) 8.46(POS) 5.98(POS)
HCV 6212-7 37 1 41 IND 8.40×104 103.00(POS) 8.22(POS) 6.56(POS)
HCV 6212-8 53 1 23 IND 2.00×105 74.53(POS) 11.20(POS) 14.80(POS)
HCV 6224-1 0 UK 13 NEG 2.30×105 0.05(NEG) 0.05(NEG) 0.03(NEG)
HCV 6224-2 3 UK 22 NEG 2.40×105 0.15(NEG) 0.05(NEG) 0.03(NEG)
HCV 6224-3 7 UK 61 NEG 1.50×105 4.05(POS) 0.06(NEG) 0.03(NEG)
HCV 6224-4 11 UK 75 NEG 8.60×104 24.77(POS) 0.15(NEG) 0.05(NEG)
HCV 6224-5 19 UK 73 IND 2.70×105 138.30(POS) 2.49(POS) 1.39(POS)
HCV 6224-6 22 UK 61 IND 3.10×105 149.10(POS) 4.11(POS) 2.19(POS)
HCV 9058-1 0 1a 19 NEG POS 0.29(NEG) 0.15(NEG) 0.14(NEG)
HCV 9058-2 3 1a 18 NEG POS 1.47(POS) 0.19(NEG) 0.20(NEG)
HCV 9058-3 7 1a 55 NEG POS 23.60(POS) 0.55(NEG) 0.50(NEG)
HCV 9058-4 10 1a 75 POS POS 88.04(POS) 1.63(POS) 0.99(GZ)
HCV 9058-5 14 1a 156 POS POS 193.00(POS) 5.13(POS) 5.18(POS)
图2 Elecsys anti-HCV Ⅱ，Architect anti-HCV和Vitros anti-HCV检测861例临床常规样本的S/Co值的分布
表2 不同抗-HCV筛查试剂检测临床常规样本的性能[%(95%CI)]
检验方法 特异度(%, 95% CI) 敏感度(%, 95% CI) PPV(%, 95% CI) NPV(%, 95% CI)
Elecsys anti-HCV Ⅱ 99.64(98.87～99.91) 100.00(78.12～100.00) 85.71(62.64～96.24) 100.00(99.43～100.00)
Architect anti-HCV 99.17(98.22～99.63) 94.44(70.63～99.71) 70.83(48.75～86.56) 99.88(99.22～99.99)
Vitros anti-HCV 99.05(98.06～99.56) 100.00(78.12～100.00) 69.23(48.10～84.91) 100.00(99.43～100.00)
图3 3种抗-HCV筛查试剂S/Co值的相关性分析(n＝17)
图4 Elecsys anti-HCV Ⅱ 和Architect anti-HCV检测临界值样本的S/Co值分布
表3 Elecsys anti-HCV Ⅱ、Architect anti-HCV筛查试剂及RIBA 3.0、Realtime HCV检测抗-HCV临界值样本的结果比较(n＝73)
例数(%) VitrosAnti-HCV ElecsysAnti-HCV Ⅱ ArchitectAnti-HCV RIBA3.0 HCV RNA(LOD 12 IU/mL) 最终诊断
11(15.07) POS NEG NEG IND NEG IND
23(31.51) POS NEG NEG NEG NT NEG
13(17.81) POS NEG POS IND NEG IND
8(10.96) POS NEG POS NEG NT NEG
1(1.37) POS POS NEG IND NEG IND
15(16.44) POS POS POS POS NT POS
2(2.74) POS POS NEG POS NT POS
表4 3种抗-HCV筛查试剂检测HIV-HCV合并感染患者的结果比较 (n＝178)
例数(%) ElecsysAnti-HCV Ⅱ ArchitectAnti-HCV VitrosAnti-HCV RIBA3.0 HCV RNA(LOD 12 IU/mL) 最终诊断
39(21.91) POS POS POS NT NEG POS
122(68.54) POS POS POS NT POS POS
11(6.18) NEG NEG NEG NT NEG NEG
3(1.69) NEG NEG NEG NT POS POSa
1(0.56) POS NEG NEG IND NEG IND
1(0.56) NEG POS NEG NEG NEG NEG
1(0.56) POS NEG POS POS NEG POS
图5 不同基因型样本HCV核心区 氨基酸序列(5-50)的比对
表5 Elecsys anti-HCV II检测不同基因型患者血清抗-HCV水平的比较
基因型 例数 年龄(岁，±s) 男/女(例) HCV RNA(log IU/mL，±s) 抗-HCV S/Co(±s)
1b 41 40.0±12.0 28/13 5.75±0.89 125.58±43.96
2a 43 43.3±14.1 28/15 5.92±0.93 121.29±44.10
3a 31 39.7±4.1 26/5 6.15±0.90 122.31±47.64
3b 43 39.8±9.2 29/14 6.06±0.83 105.53±43.26a
6a 45 36.8±9.4 27/18 6.29±0.84 122.02±38.90
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