Comparison of Versant HBV DNA 3.0 and COBAS AmpliPrep–COBAS TaqMan assays for hepatitis B DNA quantitation: Possible clinical implications

Garbuglia, Anna Rosa; Angeletti, Claudio; Lauria, F.; Zaccaro, Paola; Cocca, A.M.; Pisciotta, Marina; Solmone, Mariacarmela; Capobianchi, Maria Rosaria

doi:10.1016/j.jviromet.2007.07.005

Cited by 17 publications

(7 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Overall, the 95% CI was less than 0.5 log 10 IU/ml in all cases, using different assays: the Cobas AmpliPrep/ Cobas TaqMan, Cobas Amplicor Monitor, and Versant bDNA assays. Despite the different methodologies underlying these assays, there is a good correlation between the quantitation of HIV and HCV RNA and HBV DNA using the TaqMan, Monitor, and bDNA assays over a wide range of RNA/DNA levels (8,22). The type of assay for RNA/DNA quantitation in the current study did not affect the overall viral load levels following storage.…”

Section: Vol 49 2011 Stability Of Hcv Hiv and Hbv Under Storage 3165mentioning

confidence: 75%

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C

et al. 2011

View full text Add to dashboard Cite

The storage of biological samples may affect detection of viral nucleic acid, yet the stability of viral nucleic acid at standard laboratory storage temperatures (؊70°C and ؊20°C) has not been comprehensively assessed. Deterioration of viral RNA and DNA during storage may affect the detection of viruses, thus leading to an increased likelihood of false-negative results on diagnostic testing. The viral loads of 99 hepatitis C virus (HCV), 41 HIV, and 101 hepatitis B virus (HBV) patient samples were measured before and after storage at ؊20°C and ؊70°C for up to 9.1 years using Versant branched DNA assays, Cobas Monitor assays, and/or AmpliPrep/AmpliScreen assays. Clinical samples stored at ؊20°C for up to 1.2 years and at ؊70°C for up to 9 years showed a statistically significant difference from baseline with respect to HCV RNA titer, although this difference was not greater than 0.5 log 10 unit. The concentration of HIV RNA in clinical samples stored at ؊20°C for 2.3 years and at ؊70°C for up to 9.1 years did not differ significantly from the baseline viral load. HBV DNA-positive clinical samples stored at ؊20°C for up to 5 years and at ؊70°C for up to 4 years differed significantly in viral load. In all studies, however, the loss of viral load of HCV, HIV, or HBV in clinical samples tested after storage at ؊20°C and ؊70°C for up to 9 years ranged from 0.01 to 0.35 log 10 IU/ml and did not exceed 0.5 log 10 , which is the estimated intra-assay variation for molecular tests. Hence, the loss was considered of minimal clinical impact and adequate for the detection of HCV, HIV-1, and HBV nucleic acids using nucleic acid assays for the assessment of the infectious risk of cell, blood, and tissue donors.

show abstract

Section: Vol 49 2011 Stability Of Hcv Hiv and Hbv Under Storage 3165mentioning

confidence: 75%

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Several recent articles compared results of HBV DNA quantification between the Versant branched DNA (bDNA) assay, using hybridization with multiple regions of the genome and presumably detecting both spliced and unspliced DNA (total DNA), and other quantitative assays using primers from the S region (Abbott RealTime HBV DNA and an in-house assay) (24,38) or the Roche Cobas TaqMan HBV DNA quantitative assay assumed to use S primers (detect only unspliced HBV DNA) (2,11,25) and another in-house assay using X region primers detecting both spliced and unspliced total HBV DNA (37). For the Abbott assay, the viral load was overall lower than it was with the bDNA assay but more so in 26% of sample pairs of genotypes C and A but not E (24) and lower by 0.67 log in Taiwanese samples mostly of genotype B (38).…”

Section: Discussionmentioning

confidence: 99%

Hepatitis B Virus DNA Splicing in Lebanese Blood Donors and Genotype A to E Strains: Implications for Hepatitis B Virus DNA Quantification and Infectivity

2012

View full text Add to dashboard Cite

Hepatitis B virus (HBV) is one of the major viruses transmissible by blood that causes chronic infection in immunocompromised individuals. The study of 61 HBV carrier blood donors from Lebanon revealed multiple patterns of spliced HBV DNA. HBV DNA splicing was examined and quantified in samples of five genotypes and in seroconversion panels. The Lebanese sample median viral load was 1.5 ×10 2 IU/ml. All strains were genotype D, serotype ayw; 35 clustered as subgenotype D1 and 7 clustered as subgenotype D2. Three splice variants (SP1, SP1A, and Pol/S) were observed in 12 high-viral-load samples. Twenty samples of each genotype, A to E, were tested for the presence of HBV spliced DNA and SP1-specific splice variant. An unspliced HBV genome was dominant, but 100% of strains with a viral load of ≥10 5 copies/ml contained various proportions of spliced DNA. SP1 was detected in 56/100 (56%) samples in levels that correlated with the overall viral load. HBV DNA quantification with S (unspliced) and X (total DNA) regions provided different levels of viral load, with the difference corresponding to spliced DNA. During the highly infectious window period, the SP1 variant became detectable shortly after the hepatitis B surface antigen (HBsAg), suggesting a correlation between the initiation of splicing and the production of detectable levels of HBsAg. The quantification of HBV DNA with primers located outside and inside the spliced region might provide different estimations of viral load and differentiate between infectious and defective viral genomes. The role of splicing neoproteins in HBV replication and interaction with the host remains to be determined.

show abstract

“…It is also important to interpret assay results within the context of the assay characteristics. For example, with the increasing sensitivity of HBV DNA assays, some patients who were previously thought to be negative for HBV DNA can be shown to have low levels of viremia 6, 7. Similarly, the absolute level of ALT may be more important than its relation to the local normal range, because values in the high end of the currently accepted range are associated with a higher risk of histological damage and disease progression,8, 9 and probably should not be accepted as being fully normal in patients presumed to be carriers or immune‐tolerant.…”

Section: Initial Evaluationmentioning

confidence: 99%

Evaluation of the patient with hepatitis B #

2009

View full text Add to dashboard Cite

The initial evaluation of a patient with hepatitis B virus infection should attempt to assess the disease activity and stage in the context of the known natural history of this infection and to properly assess the needs for treatment and surveillance. In addition to a medical history and focused physical examination, the initial evaluation usually requires serological, biochemical, and virological tests to confirm the diagnosis as well as an imaging study to establish a baseline for future monitoring. A liver biopsy is generally not needed but can provide useful information on prognosis, need for surveillance for hepatocellular carcinoma (HCC), and whether to recommend therapy. Follow-up monitoring is aimed at determining disease progression, development of complications, and reassessing the need for treatment.

show abstract

Comparison of Versant HBV DNA 3.0 and COBAS AmpliPrep–COBAS TaqMan assays for hepatitis B DNA quantitation: Possible clinical implications

Cited by 17 publications

References 28 publications

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C

Hepatitis B Virus DNA Splicing in Lebanese Blood Donors and Genotype A to E Strains: Implications for Hepatitis B Virus DNA Quantification and Infectivity

Evaluation of the patient with hepatitis B #

Contact Info

Product

Resources

About

Comparison of Versant HBV DNA 3.0 and COBAS AmpliPrep–COBAS TaqMan assays for hepatitis B DNA quantitation: Possible clinical implications

Cited by 17 publications

References 28 publications

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 o C and −70 o C

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 o C and −70 o C

Hepatitis B Virus DNA Splicing in Lebanese Blood Donors and Genotype A to E Strains: Implications for Hepatitis B Virus DNA Quantification and Infectivity

Evaluation of the patient with hepatitis B #

Contact Info

Product

Resources

About

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C

Stability of Hepatitis C Virus, HIV, and Hepatitis B Virus Nucleic Acids in Plasma Samples after Long-Term Storage at −20 ^o C and −70 ^o C