HCV Real Time PCR Kit
The Hepatitis C Virus, a member of the Flaviviridae, is an enveloped virus with a single-stranded positive-sense RNA genome. The infection onset is insidious, with anorexia, vague abdominal discomfort, nausea and vomiting, progressing to jaundice (less frequently than hepatitis B). The severity ranges from unapparent cases in approximately 90% of infections to rare, fulminating, fatal cases. Chronic liver disease with fluctuating or persistently elevated liver enzymes is common, occurring after 50%-80% of adult HCV infections. Of those with chronic liver disease, 30%-60% may develop chronic active hepatitis, and 5%-20% may develop cirrhosis. Chronic infection is often not symptomatic; there appears to be an association between HCV infection and hepatocellular carcinoma; of these chronically infected persons, approximately 50% will develop cirrhosis or cancer of the liver. The virus is parenterally transmitted. Percutaneous exposure to contaminated blood (102 -103 infectious particles/mL of blood) and plasma derivatives, contaminated needles, and syringes are essential vehicles of spread, especially among injecting drug users. The risk of HCV transmission by household contact and sexual activity has not been well defined, but the efficiency of information via these routes appears to be low. A vertical message appears to be uncommon. However, risk of transmission may increase when the mother is co-infected with HIV. In over 40% of cases, the risk factor(s) for HCV transmission cannot be identified.
ViroGene HCV 1.0 Real-Time PCR Kit is based on the real-time PCR principle. The pathogen is detected using fluorescent dyes that are incorporated into oligonucleotide probes. The assay utilizes the 5’ exonuclease activity of Taq Polymerase to cleave a dual labeled fluorescent hydrolysis probe during the extension phase of PCR
Kit features
- Dual Probe Assay
The highly mutable nature of pathogens often proves challenging in molecular assay design. Traditional qPCR assays are designed so that a single primer and probe set targets one location within the pathogen genome. However, these single-target assays often prove inadequate to detect and quantify viral targets. Sequence polymorphisms in the assay target region can cause impaired primer or probe binding, which may result in significant under-quantification or a false negative result. To eliminate this possibility, ViroGene HCV RT-qPCR assay targets two regions within the pathogen genome, utilizing unique primer and probe sets for each target site. The driving principle behind our multiple-target assay design is that when a polymorphism occurs in one assay target, the additional sets of primers and probes for the alternate target region will still detect and accurately quantify the pathogen and thus reduces the likelihood of under-quantification or a false negative result.
- All kit Components are Liquid Ready to use with enhanced freeze-thaw stability (> 30 times)
- Using the highest standards for the choice of enzymes on our kits which can tolerate all types of inhibitors found usually in clinical samples with no effect on result quality
- Common Internal control across a wide range of assays.
- Common Reagents and common protocols, which ensures running more than one test type on the same run.
- Kit components enough to perform 110 Tests, to perform calibration, and QC at no extra cost.
| Item Name | HCV |
| Technology | one -step RT real-time PCR |
| Type of Analysis | qualitative and quantitative |
| Target Sequence | conservative region of 5’ UTR sequence |
| Specificity | HCV genotype 1–8, 100 % |
| Sensitivity (LoD) | reaches up to 25.0 IU/ml with probability of 95 % (on HCV NIBSC 02/264/01using manual extraction Gene Extract Viral RNA/DNA Extraction Kit) reaches up to 20 IU/ml with probability of 95 % (on HCV NIBSC 96/102 using automatic extraction MagCore NA Extractor |
| Linear Range | 1010 – 20 IU/ml with precision of ± 0.5 log |
| Reporting Units | IU/µl |
| Conversion Factor | 1 IU = 5.62 cp |
| Metrological Traceability | HCV NIBSC 14/150 |
| Extraction/Inhibition Control | Exogenous Internal Control |
| Validated Specimen * | plasma, serum |
| Storage | -20 ± 5 °C |
| Validated Extraction Method | ViroGene Viral RNA/DNA Extraction Kit & MagCore NA Extractor |
| Validated Instruments * | Applied Biosystems 7300 / 7500 Real-Time PCR System
AriaMx Real-Time PCR System CFX Connect™ / CFX96™ / Dx Real-Time PCR Detection System DTlite Real-Time PCR System LightCycler® 2.0 / 480 LineGene 9600 / 9600 Plus QuantStudio™ 3 Real-Time PCR System Rotor-Gene 3000 / 6000 / Q SLAN® Real-Time PCR System |
| Required Detection Channels | FAM, HEX |
| Certification | CE IVD |
