HBV Real Time PCR Kit

The Hepatitis B Virus, a member of the Hepadnaviridae, is an enveloped virus with a partially double-stranded DNA genome. The infection can be asymptomatic or symptomatic, which starts with anorexia, vague abdominal discomfort, nausea and vomiting, sometimes arthralgias and rash, often progressing to jaundice. Fever may be absent or mild; severity ranges from inapparent cases to fatal acute hepatic necrosis or chronic infection. Long-term fatality rate is 2-3% due to cancer or cirrhosis of the liver; 95% of adult infections are self-limited. The mode of transmission is through percutaneous or permucosal exposure to infectious body fluids (blood, blood products, cerebrospinal fluid, serum-derived fluids, saliva, semen, vaginal fluids, unfixed tissues, and organs), indirect contact with contaminated items in the laboratory; commonly spread by contaminated needles, syringes, and other equipment; contamination of wounds or lacerations; exposure of mucous membranes; sexual contact, household contact, perinatal transmission from mother to infant, nosocomial exposure

ViroGene HBV 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

• 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 HBV qPCR assay targets two regions within the pathogen genome, utilizing unique primer and probe sets for each target site( Surface & Pre Core ) . 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 choice of enzymes on our kits which can tolerates all types of inhibitors found usually in clinical samples with no effect on result quality 
• The employs a UDG enzyme to prevent carry over contamination from previous runs, which acts at Room Temperature during reaction setup with no special pre-Run incubation step. 

• 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	HBV
Technology	Real-time PCR
Type of Analysis	qualitative and quantitative
Target Sequence	DNA conservative sequence of S Region & Pre Core Region
Specificity	HBV genotype A-H, pre-core mutants HBV (HBeAg negative), 100 %
Sensitivity (LoD)	reaches up to 10 IU/ml with probability of 95 % (on HBV NIBSC 02/264/01 using manual extraction Gene Extract Viral RNA/DNA Extraction Kit)
Linear Range	1010 – 102.5 IU/ml with precision of ± 0.5 log
Dynamic Range	1010 – 36.9 IU/ml
Reporting Units	IU/µl
Conversion Factor	1 IU = 4.84 cp
Metrological Traceability	HBV NIBSC 10/264
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