Hepatitis B virus

Kits: EliGene® HBV RT Kit (ref: 90037-RT)
EliGene® HBV UNI kit (ref: 90037-UNI)
Package size:
50 reactions
Possibility of quantification:
EliDNA HBV standards (included with the kits)
Analytical specificity: Hepatitis B virus
Analytical sensitivity: 0.1 IU/ml of isolated specimen
Specimens: serum, plasma
Compatible instruments:
RT Kit intended for ABI 7000, 7300, 7500 (Applied Biosystems), LightCycler® 480 and LightCycler® Nano (Roche), RotorGene 6000 or RotorGene Q (Qiagen); MyGo Mini, MyGo Pro (IT-IS Life Science)
UNI Kit intended for LightCycler® 2.0
CE certification: CE 1023
Detected DNA region: genome (core protein)
Detection technology: TaqMan probes (RT version), Moleular Beacons (UNI version)
Clinical study description and results:
Within the frame of testing the functional characteristics of EliGene® HBV RT Kit and EliGene® HBV UNI Kit overall 500 clinical specimens were analyzed. From these specimens, 300 HBV positive specimens and 200 HBV negative specimens were confirmed by artus® HBV TM PCR Kit. The EliGene® HBV RT Kit and EliGene® HBV UNI Kit diagnosed as HBV positive 299 specimens. 1 specimen from Nigeria was determined by the EliGene® HBV RT Kit and EliGene® HBV UNI Kit as negative. The sensitivity and specificity of EliGene® HBV RT Kit and the EliGene® HBV UNI Kit is 98.78% and 100%, respectively. EliGene® HBV RT Kit and the EliGene® HBV UNI Kit shows 99,8% (499 from 500 specimens) match in detection of HBV DNA compared to results determined by artus® HBV TM PCR Kit.

100% sensitivity can be declared for both kits for the concentration of 5 IU/ml of serum corresponding to concentration of 0.1 IU/ml of isolated specimen by Magneto 1ml Serum/Plasma DNA/RNA isolation kit (see page 34) - extraction volume: 1.0 ml, elution volume: 50 μl. The linear range of the EliGene® HBV RT Kit and EliGene® HBV UNI Kit has been determined to cover concentrations from 0.1 IU/μl to at least 1 x 105 IU/μl.

Precision data of the EliGene® HBV RT Kit were calculated on basis of the Cp values of the amplification curves using the Quantification standard 4 (10 000 IU/ml) and Internal Control (Figure 7).
Pathogen description:
Hepatitis B virus is a small DNA virus divided into Hepadnaviridae family. Eight different genotypes signed by alphabetical letters A to H, have been previously described. Since the proposal of the four described genotypes (A–D), four others (E–H) have been characterized during the two last decades. Recently, a ninth “genotype” evidenced in North-West China, India, Lao and Vietnam and tentatively termed “I” was suggested, although it is still subject to debate and as being a recombinant strain with a genotype C backbone. Finally, very recently, a tenth genotype provisionally assigned to genotype “J” was proposed for a Japanese patient's HBV isolate.

Hepatitis B (HB) is transmitted by the exchange of body fluids e.g. blood, semen, breast milk and saliva. The virus spread between people who have unprotected sexual intercourse, drug users who share needles and syringes and health care workers in contact with potentially contaminated blood or body fluids.

It is estimated that one third of the world's population has serological evidence of past or present infection with HBV and that 350 to 400 million people are still chronically infected, of whom 78 % lived in Asia, 16 % in Africa, 3 % in South America and the remaining 3% in Europe, North America and Oceania.

HBV infection has a broad spectrum of clinical diseases, ranging from acute hepatitis (including fulminant hepatic failure) to a low viraemic asymptomatic “inactive” carrier state or to progressive chronic hepatitis that may lead to cirrhosis with an annual rate of 2 to 5 % in HBe-positive patients and hepatocellular carcinoma (HCC) with a cumulative 5-year incidence of 15 to 20 %. Both HBV-related end-stage liver disease and HCC are responsible for around 1 million deaths per year.
References:
Bannister BA, Begg NT, Gillespie SH. 2000. Infectious Disease. Blackwell Science, 2th Ed.

Ho SKN, Yam W-C, Leung ETK, Wong L-P, Leung JKH, Lai K-N, Chan TM. 2003. Rapid quantification of hepatitis B virus DNA by real-time PCR using fluorescent hybridization probes. J Med Microbiol. 52(5): 397-402.