Laboratory Evaluation of a Fully Automated Chemiluminescence Immunoassay for Rapid Detection of HBsAg, Antibodies to HBsAg, and Antibodies to Hepatitis C Virus

Ismail, Nahed; Fish, Geoffrey; Smith, Michael B.

doi:10.1128/jcm.42.2.610-617.2004

Cited by 49 publications

(34 citation statements)

References 51 publications

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“…CLIA showed significantly improved specificity, a greater positive predictive value, and a similar sensitivity compared to those of EIA for detecting anti-HCV antibodies (10, 15). Although automated CLIAs are gradually replacing the EIA, there are no published studies on the comparative evaluation of automated CLIAs (10,15,16,22,27). We compared the performance of currently marketed anti-HCV automated CLIAs under routine conditions of a hospital laboratory.…”

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confidence: 99%

Clinical Performance Evaluation of Four Automated Chemiluminescence Immunoassays for Hepatitis C Virus Antibody Detection

Kim

Yoon

et al. 2008

J Clin Microbiol

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Various automated chemiluminescence immunoassay (CLIA) analyzers for the detection of antibodies to hepatitis C virus (HCV) are now commercially available in clinical laboratories and are replacing conventional enzyme immunoassays. We investigated the performance of four anti-HCV CLIAs (the Architect Anti-HCV assay on the Architect i2000 system, the Vitros Anti-HCV assay on the Vitros ECiQ Immunodiagnostic System, the Access HCV Ab PLUS assay on the UniCel DxI 800 analyzer, and the newly developed Elecsys Anti-HCV assay on the Cobas e 411 analyzer). The total percent coefficient of variation values of imprecision were 3.5 to 5.7% with positive control materials and 7.2 to 10.2% with negative control materials. The agreement between the results of the Elecsys, Architect, Vitros, and Access CLIAs ranged from 94.5 to 98.1%. The clinical sensitivity of all CLIAs was 100%. Each CLIA showed excellent reproducibility and clinical sensitivity. The Elecsys, Architect, Vitros, and Access CLIAs showed clinical specificities of 98.2, 98.8, 96.5, and 98.2%.Hepatitis C virus (HCV), first identified in 1989, is an enveloped positive-strand RNA virus classified in the Hepacivirus genus in the family Flaviviridae (6). The HCV genome is about 9.5 kb in length and encodes 3,011-to 3,033-amino-acid polypeptides in structural and nonstructural regions (20). The structural region contains the core protein and two envelope proteins (E1 and E2), and nonstructural proteins have been assigned protease (NS2, NS3, and NS4A), helicase (NS3), and RNA-dependent RNA polymerase (NS5B) (21) functions.The first commercially available anti-HCV enzyme immunoassay (EIA) used a single HCV recombinant antigen derived from the nonstructural NS4 protein designated c100-3 (19). The sensitivity of this first-generation EIA was low for a highprevalence population (approximately 80%) and showed a high false-positive rate (up to 70%) in a low-prevalence blood donor group (13). Therefore, a second-generation EIA was developed and approved for use by the Food and Drug Administration (FDA) in 1992 (3). The second-generation EIA, which contained additional HCV antigens from the core (c22-3) and NS3 (c33c) proteins, showed increased sensitivity and specificity and shortened the average seroconversion period from 16 to 10 weeks (1, 3, 13, 18). The third-generation EIA, which added a fourth antigen (NS5), showed significantly improved performance, particularly for high-risk patients (2, 8). However, a residual risk still exists due to the seroconversion period of approximately 56 days, and high false-positive rates were not resolved (12). The Centers for Disease Control and Prevention (CDC) recommended that an anti-HCV screening test positive result be verified by a more specific supplemental assay such as recombinant immunoblot or nucleic acid test (5). To facilitate the use of the supplemental assay, the revised guideline included an option for reflex supplemental testing based on signal-to-cutoff (s/co) ratios (4).Today, automated chemiluminescence immunoassay (...

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Clinical Performance Evaluation of Four Automated Chemiluminescence Immunoassays for Hepatitis C Virus Antibody Detection

Kim

Yoon

et al. 2008

J Clin Microbiol

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“…The VITROS anti-HCV CIA has recently been shown to be at least as specific and sensitive as conventional EIAs (7,9), and use of this CIA has been increasing. However, detailed studies of the CIA S/Co ratio value predictive of a positive supplemental HCV test have been limited (7,9).…”

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confidence: 99%

“…However, detailed studies of the CIA S/Co ratio value predictive of a positive supplemental HCV test have been limited (7,9). The CDC guidelines have suggested reflex supplemental testing for samples with S/C ratios of Ͻ8.0 in the VITROS anti-HCV assay based on the evaluation of a total of 1,326 reactive samples, with only 142 of these specimens having S/Co ratios of Յ7.9 (2).…”

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confidence: 99%

“…Ismail et al compared the performance of the VITROS Anti HCV assay to the Abbott second-generation EIA; 318 HCV-negative samples and 177 HCV-positive samples in the Abbott assay were evaluated, but confirmatory testing was only performed on the eleven discrepant results (9). Among the seven VITROS anti-HCV-positive samples, RIBA was negative in four samples and indeterminate in three samples; all seven samples had S/Co ratios of Ͼ1 and Ͻ8, but the true status of the three indeterminate samples was not determined.…”

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Efficiency of the Ortho VITROS Assay for Detection of Hepatitis C Virus-Specific Antibodies Increased by Elimination of Supplemental Testing of Samples with Very Low Sample-to-Cutoff Ratios

et al. 2005

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The clinical significance of specimens with low sample-to-cutoff (S/Co) ratios in the Ortho VITROS chemiluminescence assay (CIA) for detection of antibodies to hepatitis C virus (HCV) was evaluated. In one study of 482 CIA-reactive samples, none of the 83 samples with S/Co ratios of <5 was HCV RNA positive. In a subsequent study, 332 samples with S/Co ratios of between 1 and 20 were tested with the recombinant immunoblot assay (RIBA). None of the 163 samples with S/Co ratios of <5 was RIBA positive, 83% were RIBA negative, and 28 samples (18%) were RIBA indeterminate. HCV RNA and/or clinical evidence of hepatitis was not found in the 27 indeterminate cases examined. These results show that over 99% of samples with very low S/Co ratios (<5) have no evidence of HCV infection. Therefore, we suggest that the HCV antibody testing algorithm for the VITROS assay might be modified to eliminate supplemental testing of samples with very low S/Co ratios.Assays for the detection of hepatitis C virus (HCV) antibodies have high false-positive rates, particularly in low-prevalence populations. Therefore, other tests such as recombinant immunoblot assay (RIBA) or HCV RNA PCR are used to confirm positive HCV antibody screening tests. To facilitate usage of the supplemental testing, the Centers for Disease Control and Prevention (CDC) published guidelines that incorporate anti-HCV assay signal-to-cutoff (S/Co) ratios into reflex testing algorithms for HCV antibody testing, in order to provide a more systematic approach for the laboratory diagnosis of HCV and minimize the number of specimens that require supplemental testing (2). Data obtained with three anti-HCV screening assays were used for these recommendations: two enzyme immunoassays (EIAs) (Abbott HCV EIA 2.0 and ORTHO HCV version 3.0 enzyme-linked immunosorbent assay) and one enhanced chemiluminescence immunoassay (CIA) (VITROS anti-HCV assay; Ortho-Clinical Diagnostics).The VITROS anti-HCV CIA has recently been shown to be at least as specific and sensitive as conventional EIAs (7, 9), and use of this CIA has been increasing. However, detailed studies of the CIA S/Co ratio value predictive of a positive supplemental HCV test have been limited (7, 9). The CDC guidelines have suggested reflex supplemental testing for samples with S/C ratios of Ͻ8.0 in the VITROS anti-HCV assay based on the evaluation of a total of 1,326 reactive samples, with only 142 of these specimens having S/Co ratios of Յ7.9 (2).We started using the VITROS anti-HCV assay shortly after its Food and Drug Administration approval in August 2001 and have prospectively applied reflex supplemental testing to all samples with S/Co ratios of between 1.00 and 20.0 in the setting of a population of U.S. veterans (8). The objectives of the present study were (i) to determine the S/Co ratio predictive of detectable HCV RNA; (ii) to evaluate, in samples with low S/Co ratios (between 1 and 20), the relationship between S/Co ratios and the number and types of bands detected by RIBA; (iii) to assess, for samples with i...

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“…Also, HBV detection methods differ in developed and developing countries according to the level of endemic HBV infections. Besides, automated HBsAg detection immunoassay systems have been developed and widely used [6][7][8][9]. However, high specificity enzyme-linked immunosorbent assays (ELISAs) are still commonly used [4,5,10,11].…”

Section: Article; Medical Biotechnology (Mb)mentioning

confidence: 99%

Cloning of anti-HBsAg single-chain variable fragments from hybridoma cells for one-step ELISA

Erdağ

Balcıoğlu

Bahadır

et al. 2017

Biotechnology & Biotechnological Equipment

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Hepatitis B virus (HBV) infection is a worldwide health problem. More than 400 million people are chronic HBV carriers in the world. Infected individuals are at a high risk of developing liver cirrhosis and hepatocellular carcinoma as the main consequences of HBV. The discoveries of fast diagnostic systems and new therapeutic applications are crucial in the fight against viral hepatitis. In this paper we present the generation of a single-chain variable fragment (scFv) from a mouse monoclonal antibody specific to the HBV surface antigen (HBsAg) and demonstrate its expression as a bacterial alkaline phosphatase (AP) fusion protein. In this study, we constructed scFvs from hybridoma cells expressing HBsAg-specific antibody using phage display technology and expressed them in Escherichia coli. The anti-HBsAg scFvs were inserted into pQE-2 vector to produce scFv antibody genetically fused to bacterial AP. Reproducibility of the recombinant HBsAg-scFv fusion protein was tested using Enzyme-linked Immunosorbent Assay (ELISA). Present preliminary findings indicate that the anti-HBsAg-scFv AP conjugate could be further used for the development of one-step ELISA for the detection of HBV.

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Laboratory Evaluation of a Fully Automated Chemiluminescence Immunoassay for Rapid Detection of HBsAg, Antibodies to HBsAg, and Antibodies to Hepatitis C Virus

Cited by 49 publications

References 51 publications

Clinical Performance Evaluation of Four Automated Chemiluminescence Immunoassays for Hepatitis C Virus Antibody Detection

Clinical Performance Evaluation of Four Automated Chemiluminescence Immunoassays for Hepatitis C Virus Antibody Detection

Efficiency of the Ortho VITROS Assay for Detection of Hepatitis C Virus-Specific Antibodies Increased by Elimination of Supplemental Testing of Samples with Very Low Sample-to-Cutoff Ratios

Cloning of anti-HBsAg single-chain variable fragments from hybridoma cells for one-step ELISA

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