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RealTime HCV

Put precision into practice.


Abbott RealTime HCV

Precise performance at
the point of decision

 

For In Vitro Diagnostic Use

The Abbott RealTime HCV assay is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for use with the Abbott mSample Preparation System reagents and with the Abbott m2000sp and m2000rt instruments for the quantitation of hepatitis C viral (HCV) RNA in human serum or plasma (EDTA) from HCV-infected individuals. Specimens containing HCV genotypes 1 – 6 have been validated for quantitation in the assay.

The Abbott RealTime HCV assay is intended for use as an aid in the management of HCV-infected patients undergoing antiviral therapy. The assay measures HCV RNA levels at baseline and during treatment and can be utilized to predict sustained and non-sustained virological response to HCV therapy. The results from the RealTime HCV assay must be interpreted within the context of all relevant clinical and laboratory findings.

Assay performance characteristics have been established for individuals treated with peginterferon alfa-2a or 2b plus ribavirin. No information is available on the assay’s predictive value when other therapies are used. Assay performance for determining the state of HCV infection has not been established.

The Abbott RealTime HCV assay is not for screening blood, plasma, serum or tissue donors for HCV, or to be used as a diagnostic test to confirm the presence of HCV infection.

Abbott RealTime HCV Performance*
Limit of Detection (LOD)
12 IU/mL
Limit of Quantitation (LOQ) 12 IU/mL
Linear Range 12 IU/mL (1.08 log IU/mL) to 100 million IU/mL (8.0 log IU/mL)
Target Region 5' utr
Genotype Detection Accurate quantitation of genotypes 1, 2, 3, 4, 5 and 6
Internal Control Added to lysis buffer during extraction and detected with all samples
Calibration External two point
Specimen Type Human serum and plasma (EDTA)
Input Volume 0.5mL
Precision SD: less than or equal 0.25 log IU/mL
Standardization Second WHO international standard to HCV RNA (NIBSC 96/798)

Abbott Realtime

An assay desgined with leading science.

The unique combinaiton of Abbott RealTime HCV assay design features ensures reliable detection of all HCV genotypes with a high degree of precision.

Abbott RealTime HCV Assay

Converved Target Region

Primers and Probe are Targeted to the Integrase Region of the Polymerase Gene

The target sequence for the Abbott RealTime HCV assay is in the 5´UTR region of the HCV genome. This region is specific for HCV and is highly conserved. The primers are designed to hybridize to the 5´UTR region with the fewest possible mismatches among HCV genotypes 1-6.

Unique Probe Design

Single Stranded Linear Probe Design

In the absence of target, the probe achieves quenching through random coiling.

Single Stranded Linear Probe Design

In presence of target, the probe prefers to hybridize to the target sequence, allowing fluorescent detection; cleavage is not required.

Optimal Cycling Conditions

Cycling Conditions: Low Temperature Read Cycles

The cycling conditions for Abbott RealTime HCV encompass a low temperature read cycle. This read cycle allows the probe to tolerate mismatches more effectively than a probe required to bind during the extension phase.

Precise External Calibration

The external calibration curve is a key assay design feature of the RealTime HCV assay to enable high precision: The use of different primers for HCV target and Internal Control (heterologous) minimizes competitive effects in the PCR reaction. The stored calibration curve reduces the variability of the viral load calculation compared to an internal calibration design.

Abbott RealTime HCV precisely quantitiates HCV levels
at clinical decision points for new DAA triple therapies

Abbott RealTime HCV Precise External Calibration

References

  1. Abbott RealTime HCV package insert (51-602124)
  2. Pisani et al. Quantification of Hepatitis C Virus (HCV) RNA in a Multicenter Study: Implications for Management of HCV Genotype 1-Infected Patients. J Clin Microbio. (2009) 47:2931-2936 
  3. Baylis et al. World Health Organization (WHO) collaborative study to calibrate the 3rd International Standard (IS) for Hepatitis C virus (HCV) RNA nucleic acid amplification technology (NAT)-based assays. Vox Sanguinis (2011) 100:409-417 
  4. Braun et al. Poster #25. ESCV 2012. Comparative analysis of viral load quantification in clinical samples with low detectable HCV viremia 
  5. Lucic et al. Poster #24. ESCV 2012. Importance of HCV viral load assay precision on the treatment of patients with Direct Acting Antivirals: Potential Impact on Response Guided Therapy (RGT) 
  6. Fevery et al. Podium presentation. EASL 2012. Comparison of two quantitative HCV RNA assays in samples from patients treated with a protein inhibitor-based therapy: implications for response-guided therapy 
  7. Cloherty et al. Poster #765. AASLD 2012. HCV RNA Assay Sensitivity Impacts the Management of Patients Taking DAA
 

Intended Use

The Abbott RealTime HCV assay is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for use with the Abbott mSample Preparation System reagents and with the Abbott m2000sp and m2000rt instruments for the quantitation of hepatitis C viral (HCV) RNA in human serum or plasma (EDTA) from HCV-infected individuals. Specimens containing HCV genotypes 1 – 6 have been validated for quantitation in the assay.

The Abbott RealTime HCV assay is intended for use as an aid in the management of HCV-infected patients undergoing antiviral therapy. The assay measures HCV RNA levels at baseline and during treatment and can be utilized to predict sustained and non-sustained virological response to HCV therapy. The results from the RealTime HCV assay must be interpreted within the context of all relevant clinical and laboratoryfindings.

Assay performance characteristics have been established for individuals treated with peginterferon alfa-2a or 2b plus ribavirin. No information is available on the assay’s predictive value when other therapies are used. Assay performance for determining the state of HCV infection has not been established.

The Abbott RealTime HCV assay is not for screening blood, plasma, serum or tissue donors for HCV, or to be used as a diagnostic test to confirm the presence of HCV infection.

 

LIMITATIONS OF THE PROCEDURE

  • Human serum and plasma specimens (EDTA) may be used with the Abbott RealTime HCV assay. The use of other anticoagulants has not been validated with the Abbott RealTime HCV assay
  • Though rare, mutations within the highly conserved regions of the viral genome covered by the Abbott RealTime HCV assay primers and/or probe may result in the under-quantitation of or a failure to detect the presence of the virus in this circumstance.
  • A specimen with a result of “Not Detected” cannot be presumed to be negative
  • for HCV RNA.
  • Precision was established with HCV Genotypes 1 and 3 only.
  • As with any diagnostic test, results from the Abbott RealTime HCV assay should be interpreted in conjunction with other clinical and laboratory findings.
  • Due to inherent differences between technologies, it is recommended that, prior to switching from one technology to the next, users perform method correlation studies in their laboratory to quantify technology differences.

WARNINGS AND PRECAUTIONS

The Abbott RealTime HCV assay is not for screening blood, plasma, serum or tissue donors for HCV, or to be used as a diagnostic test to confirm the presence of HCV infection.

 

CAUTION:
United States Federal law restricts this device to sale and distribution to or on the order of a physician or to a clinical laboratory; and use is restricted to, by, or on the order of a physician.

For In Vitro Diagnostic Use