Sensitive and unique qPCR assay for quantifying HCV viral load.
- Sensitive quantitation of HCV viral load
- Low-cost alternative to other methods of viral load detection
- Compatible with most open qPCR platforms
- Complete set of controls including: quantitation controls, an internal sample extraction control, and a negative control
- Smaller sample input allows remaining extracted samples to be used in other tests
The AMPIPROBE® HCV assay is compatible with any properly calibrated qPCR thermal cycler capable of exciting and reading emissions from FAM/Green and CY5/Red. It has been validated for use on the following instruments: QIAGEN Rotor-Gene Q and Applied Biosystems® 7500.
The AMPIPROBE® HCV assay kit is a quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay for the quantitative detection of human hepatitis C virus (HCV) RNA in plasma or serum. The proprietary primer mix included in the kit is specific for HCV genotypes 1 through 6. The kit includes titered high, medium, and low HCV controls which enable virus quantitation in IU/mL. In addition, the kit contains an extraction control to ensure proper sample nucleic acid extraction.
Enzo's AMPIPROBE® technology incorporates probe detection technology in primer design. It employs a combination of fluorescent reporter-labeled primers and quencher-labeled primers to amplify DNA, akin to traditional PCR. When free in solution, fluorescent primers generate a signal. However, as the primers are incorporated into amplified DNA, the quencher and the fluorophore are brought within close proximity and exhibit Forster resonance energy transfer (FRET). This causes a logarithmic decay of signal with respect to the number of amplification cycles of DNA. Once the signal decays to a defined threshold, a value is generated with respect to the corresponding cycle. The threshold cycle is indicative of the amount of target RNA or DNA in the sample.
In a validation study approved by the New York State Department of Health, the AMPIPROBE® HCV assay was determined to have the following sensitivity:
LOD serum - 5.46 IU/mL via 95% Probit analysis
LOD plasma - 7.91 IU/mL via 95% Probit analysis
LOQ serum - 10 IU/mL via 95% hit rate analysis
LOQ plasma - 10 IU/mL via 95% hit rate analysis.
In the same validation study, the linear range of both serum and plasma was determined to be 5 to 25,000,000 IU/mL.
