Assessment of the Target-Capture PCR Hepatitis B Virus (HBV) DNA Quantitative Assay and Comparison with Commercial HBV DNA Quantitative Assays

Arcangel, Phillip; Cottrell, Joshua; Coit, Doris; Medina-Selby, Angelica; McCoin, Colin S.; Madriaga, Dennis; Chien, David; Phelps, Bruce H.

doi:10.1128/jcm.42.11.5199-5204.2004

Cited by 26 publications

(18 citation statements)

References 26 publications

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“…However, accurate conversion factors are likely to be dependent on the chemistry used for HBV DNA quantification. In support of this, PCR-based quantification assays were demonstrated to have similar conversion factors (Amplicor, 7.3 copies per IU; SuperQuant, 6.2 copies per IU) that were higher than those for bDNA (VERSANT, v2.0, 4.5 copies per IU) and Hybrid Capture II (2.3 copies per IU) (132).…”

Section: Molecular Tests For Hbv Quantification: Available Assays Andmentioning

confidence: 92%

Molecular Testing in the Diagnosis and Management of Chronic Hepatitis B

Valsamakis

2007

Clin Microbiol Rev

135

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SUMMARY Hepatitis B virus (HBV) is an enveloped virus with a small (3.2-kb) partially double-stranded DNA genome that causes acute and chronic infections. The impact of these infections on public health worldwide is enormous, with an estimated prevalence of 2 billion acute infections and 360 million chronic infections globally. This review focuses on chronic hepatitis B and the molecular assays used in its diagnosis and management. Background information, including that about features of the hepatitis B virion, viral replication, and epidemiology of infection, that is important for understanding chronic hepatitis B and molecular diagnostic tests for HBV is provided. To facilitate an understanding of the utility of molecular testing for chronic hepatitis B, the four stages of chronic hepatitis B infection that are currently recognized, as well as an additional entity, occult hepatitis B, that can be diagnosed only by sensitive nucleic acid amplification methods, are reviewed in detail, including available therapeutic agents. The molecular diagnostic content focuses on tests for HBV DNA quantification, genotyping, and mutation detection (including precore/core promoter and antiviral resistance mutations). The discussion of these tests encompasses their current utility and performance characteristics, drawing from current clinical guidelines and other studies from the literature. In recognition of the continual evolution of this field, the final section describes emerging molecular markers with future diagnostic potential.

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Section: Molecular Tests For Hbv Quantification: Available Assays Andmentioning

confidence: 92%

Molecular Testing in the Diagnosis and Management of Chronic Hepatitis B

Valsamakis

2007

Clin Microbiol Rev

135

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“…Our in-house HBV PCR results are expressed in HBV IU/ml because we used a commercially available reference material calibrated against the international standard to generate a standard curve. The expected HBV results were expressed in HBV copies/ml and were divided by 5 (Ϫ 0.70 log HBV IU/ml) (14) to obtain the reference results expressed in HBV IU/ml.…”

Section: Resultsmentioning

confidence: 99%

Automated Extraction of Viral-Pathogen RNA and DNA for High-Throughput Quantitative Real-Time PCR

2005

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The performance of the m1000 system (Abbott Laboratories, Illinois) as a front-end extraction system for high-throughput "in-house" quantitative real-time PCR assays was analyzed and compared to that of manual extraction of plasma and serum samples ( hepatitis C virus [HCV] and hepatitis B virus [HBV]) and EDTAblood samples (cytomegalovirus [CMV] and Epstein-Barr virus [EBV]). Linearity of extraction was tested on dilution series of HCV and HBV reference materials. The correlation coefficient for standard curves based on repeated extraction runs was 0.97 ؎ 0.06 for HCV and 0.97 ؎ 0.03 for HBV, indicating a linear extraction from 100 to 1.0 ؋ 10 5 HCV IU/ml and from 100 to 1.0 ؋ 10 6 HBV IU/ml. Intra-and interrun variability was below 0.23 log 10 IU/ml for 2.98 to 5.28 log 10 HCV IU/ml and 2.70 to 5.20 log 10 HBV IU/ml. Correlation between automated and manual extraction was very good. For HCV, the correlation coefficient was 0.91 and the mean difference in viral load was 0.13 log 10 HCV IU/ml. For HBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.61 log 10 HBV IU/ml. For CMV and EBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.33 log 10 copies/ml. Accuracy was confirmed with a reference panel (QCMD, Glasgow, Scotland) for all four assays. No cross-contamination was observed when extracting strongly positive polyomavirus samples (8.10 log 10 copies/ml) interspersed with polyomavirus-negative samples. Automated extraction via the m1000 system offers a high reliability of extraction and resulted in a strong reduction of the required extraction hands-on time for high-throughput PCR compared to manual extraction protocols.Real-time thermocyclers have greatly decreased the amount of hands-on time in nucleic acid (NA)-based diagnostics for pathogens. The use of commercially available universal master mixes that contain all reagents except the pathogen-specific primer-probe combination has further decreased the number of pipetting steps and thus reduced labor and the possibility of errors.NA extraction has now become the most critical and laborintensive step in NA-based diagnostic assays. The overall sensitivity of the assay is determined by the NA yield, its purity, and the amount of sample equivalents that can be transferred into the amplification reaction. Conventional manual sample preparation methods are labor intensive and susceptible to contamination, handling variations, or errors (3, 9, 12).Since both the pathogen range and the number of different sample types are expanding and since multiplex downstream testing is becoming standard practice, there is a need for a generic extraction method (2, 5, 10). Ideally, the NA extraction procedure should yield pure NA from different pathogens and from a broad range of sample types.Recently, Abbott introduced m1000, an automated generic RNA and DNA extraction system using magnetic microparticle processing (17, 18). The m1000 system provides the necessary features for full automation of NA extraction of up to...

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“…Serial dilutions of the NICPBP standard (1 × 10 5 , 1 × 10 4 , 1 × 10 3 , 1 × 10 2 IU/ml) were tested in triplicate by RtF-LAMP (Table 1). The conversion factor for the results obtained was 4.4 copies/IU, and was comparable to 7.3 by the Amplicor test, 6.2 by SuperQuant, 4.5 by Quantiplex, and 2.3 by Hybrid Capture test (Shyamala et al, 2004).…”

Section: Conversion Factor Of Hbv Dna Unitsmentioning

confidence: 98%