RealTime HBV
- Indications and Limitations of Use
- Package Insert For Information Only - Not a Controlled Copy
| Product Name | Quantity | Order # | Additional Information | ||
| Abbott RealTime HBV Calibrator Kit | 12 Cal A, 12 Cal B |
02N40-70 |
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| Abbott RealTime HBV Control Kit | 8 Low Positive, |
02N40-80 |
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| Abbott RealTime HBV Amplification Reagent Kit | 96 Assays |
02N40-90 |
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| Abbott RealTime HBV Application CD-ROM | 1 each | 02N43 |
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Abbott RealTime HBV assay is an in vitro polymerase chain reaction (PCR) assay for use with the Abbott m2000 system DNA reagents and with the Abbott m2000sp and m2000rt instruments for the quantitation of Hepatitis B Virus (HBV) DNA in human serum or plasma (EDTA) from chronically HBV-infected individuals. The assay is intended for use as an aid in the management of patients with chronic HBV infection undergoing anti-viral therapy. The assay can be used to measure HBV DNA levels at baseline and during treatment to aid in assessing response to treatment. The results from the Abbott RealTime HBV assay must be interpreted within the context of all relevant clinical and laboratory findings. Assay performance for determining the clinical stage of HBV infection has not been established. Clinical performance characteristics have been established for individuals treated with adefovir diprivoxil. This assay is not intended for use as a screening test in blood or blood products for HBV or as a diagnostic test to confirm the presence of HBV infection.
| Abbott RealTime HBV Performance* |
|
| Sensitivity |
10 IU/mL for 0.5 mL sample volume 15 IU/mL for 0.2 mL sample volume |
| Linear Range |
10 IU/mL (1.00 log IU/mL) to 1 billion IU/mL (9.00 log IU/mL) |
| Precision1 |
Standard Deviation ≤ 0.25 log IU/mL |
| Specificity2 |
100% |
| Genotype Detection |
Accurate quantitation of genotypes A, B, C, D, E, F, G, H |
| Specimen Type |
Serum and Plasma (EDTA) |
| Standardization |
World Health Organization (WHO) International Standard for Hepatitis B Virus DNA (97/746) |
| Internal Control |
Added to lysis buffer during extraction and detected at all levels |
*Performance data shown from internal verification.
1 Total precision includes within-run, between-run and between-lot/instrument components of precision.
2 The specificity of the RealTime HBV assay was evaluated by testing 124 HBV seronegative serum and 125 HBV seronegative plasma specimens. HBV DNA was not detected, resulting in 100% (249/249) specificity.
| Detection and Quantitation of HBV Genotypes |
Genotype dilution linearity was demonstrated by diluting eight specimens, one of each genotype A through H, to target concentrations of 4.47 log IU/mL, 3.47 log IU/mL, 2.47 log IU/mL, and 1.17 log IU/mL. Three replicates were tested at each concentration for each genotype. The RealTime HBV assay is capable to quantitate different HBV genotypes across linear range with deviation of not more than 0.51 log IU/mL.
| Linear Range |
The upper limit of quantitation (ULQ) for the Abbott RealTime HBV assay is 1 billion IU/mL and the lower limit of quantitation is equivalent to LOD (10 IU/mL for the 0.5 mL sample preparation procedure, 15 IU/mL for the 0.2 mL sample preparation procedure).
A 13-member panel prepared by diluting an HBV positive specimen targeted from 9.13 log IU/mL to 0.29 log IU/mL in HBV negative human plasma was tested.
The RealTime HBV assay was shown to be linear in plasma across the range of HBV DNA concentrations.
Development Philosophy
Today's clinical molecular diagnostics laboratory must have confidence in the quality of HBV patient results. As a result of our real-time PCR development philosophy, Abbott RealTime HBV is engineered to quantify all HBV genotypes A-H.
Critical Target Region Selection
The selection of a highly conserved region within the Surface gene provides for the accurate detection of genotypes A-H.
The location of the target region in the N-terminal third of the Surface gene ensures that the assay is not impacted by YMDD mutants, HBsAg escape mutants, or drug resistant mutants, as this region is essential for the assembly and secretion of subviral particles, and tolerates only minor structural changes.
Hepatitis B Virus Genome and Target Region
Primers and probe are targeted within an essential segment of the Surface gene.
INTENDED USE
Abbott RealTime HBV assay is an in vitro polymerase chain reaction (PCR) assay for use with the Abbott m2000 SystemDNA reagents and with the Abbott m2000sp and m2000rt instruments for the quantitation of Hepatitis B Virus (HBV) DNA in human serum or plasma (EDTA) from chronically HBV-infected individuals. The assay is intended for use as an aid in the management of patients with chronic HBV infection undergoing anti-viral therapy. The assay can be used to measure HBV DNA levels at baseline and during treatment to aid in assessing response to treatment. The results from the Abbott RealTime HBV assay must be interpreted within the context of all relevant clinical and laboratory findings. Assay performance for determining the clinical stage of HBV infection has not been established. Clinical performance characteristics have been established for individuals treated with adefovir dipivoxil. This assay is not intended for use as a screening test in blood or blood products for HBV or as a diagnostic test to confirm the presence of HBV infection.
LIMITATIONS OF THE PROCEDURE
• Human serum and plasma (EDTA) may be used with the Abbott RealTime HBV assay. The use of other anticoagulants has not been validated for use with the Abbott RealTime HBV assay.
• The use of specimens collected in serum tubes that contain Z-clot activator, or similar types of rapid clot activator, may cause inhibited results in the RealTime HBV assay.
• In rare cases, a very low level positive result may occur from cross contamination during processing of an adjacent specimen with an extremely high copy number. Carryover rates in representative studies ranged from 0% to 2%. Per treatment guidelines, a 1 log increase is needed in order to impact patient management. In addition, treatment guidelines require two consecutive elevated measurements to occur before changing patient management.
• A single Entecavir mutation (rtA97V) occurs within the reverse primer. Of all known resistance mutations, it is the only one that occurs within any Abbott RealTime HBV primer or probe sequence. Software simulation predicts that this mutation (rtA97V) would not be expected to interfere with assay results when using RealTime HBV assay conditions.
• A specimen with a result of “Not Detected” cannot be presumed to be negative for HBV DNA.
• Drug interference was evaluated using a plasma matrix, and was not evaluated in serum. The listed drugs were tested in pools, and individual drug effects were not assessed.
• The interference studies were performed with an HBV DNA concentration of 2,933 IU/mL (3.47 log IU/mL). Potential interference on HBV DNA concentrations close to the assay LLQ (LLoQ) was not assessed.
• Results from the Abbott RealTime HBV assay should be interpreted in conjunction with other clinical and laboratory findings.
WARNINGS AND PRECAUTIONS
The Abbott RealTime HBV assay is not intended for use in the screening of blood, plasma, or tissue donors for HBV, or to be used as a diagnostic test to confirm the presence of HBV infection.
