Hi  Shaifali Gupta,

You might find all information you requested in the following paper of T. Welzel et al. “Real-Time PCR Assay for Detection and Quantification of Hepatitis B Virus Genotypes A to G”, published in J Clin Microbiol, v.44(9); 2006 PMC1594718

ABSTRACT: The detection and quantification of hepatitis B virus (HBV) DNA play an important role in diagnosing and monitoring HBV infection as well as assessing therapeutic response. The great variability among HBV genotypes and the enormous range of clinical HBV DNA levels present challenges for PCR-based amplification techniques. In this study, we describe the development, evaluation, and validation of a novel real-time PCR assay designed to provide accurate quantification of DNA from all eight HBV genotypes in patient plasma specimens. A computer algorithm was used to design degenerate real-time PCR primers and probes based upon a large number (n = 340) of full-length genomic sequences including HBV genotypes A to H from Europe, Africa, Asia, and North and South America. Genotype performance was tested and confirmed using 59 genotype A to G specimens from two commercially available worldwide genotype panels. This assay has a dynamic range of at least 8 log10 without the need for specimen dilution, good clinical intra- and interassay precision, and excellent correlation with the Bayer Diagnostics VERSANT HBV DNA 3.0 (branched DNA) assay (r = 0.93). Probit analysis determined the 95% detection level was 56 IU/ml, corresponding to 11 copies per PCR well. The high sensitivity, wide linear range, good reproducibility, and genotype inclusivity, combined with a small sample volume requirement and low cost, make this novel quantitative HBV real-time PCR assay particularly well suited for application to large clinical and epidemiological studies.

Best wishes,

Ilya Tsyrlov

Dear Shaifali,

I recommend you to go for the 3rd WHO International Standard for Hepatitis B Virus for Nucleic Acid Amplification Techniques (NIBSC code: 10/264). You can refer to the links and references provided below for more detailed information on this standard and NIBSC website for purchase enquiry.

SD

Heermann KH, Gerlich WH, Chudy M, Schaefer S, Thomssen R.

Quantitative detection of hepatitis B virus DNA in two international reference

plasma preparations. Eurohep Pathobiology Group. J Clin Microbiol.

1999;37:68-73.

Fryer JF, Heath AB, Wilkinson DE, Minor PD and the collaborative study

group. Collaborative study to evaluate the proposed 3rd WHO International

Standard for hepatitis B virus (HBV) for nucleic acid amplification technology

(NAT)-based assays. WHO ECBS Report 2011; WHO/BS/2011.2170.

http://www.nibsc.org/

http://www.nibsc.org/documents/ifu/10-264.pdf

Thank u sir...

See these links may be useful for you.

1) Multilaboratory Evaluation of Real-Time PCR Tests for ...

 www.ncbi.nlm.nih.gov › ... › PubMed Central (PMC) Traduire cette page

de AM Caliendo - ‎2011 - ‎Cité 30 fois - ‎Autres articles Multilaboratory Evaluation of Real-Time PCR Tests for Hepatitis B Virus DNA Quantification ... False-positive results were observed with all assays; the highest rates occurred ... An internal control (IC) for the RealTime and artus HBV assays and a ... Overall, there was good agreement in viral load values between the assays.

2) Comparison of the AdvanSure HBV Real-time PCR Test ...

 www.annclinlabsci.org/content/43/2/230.full.pdf - Traduire cette page de H Kim - ‎2013 - ‎Cité 6 fois - ‎Autres articles HBV PCR showed good correlations with the three other HBV DNA assays. ... Key words: hepatitis B virus; DNA quantification; real-time PCR; AdvanSure .... positive control; NC, negative control; LP, low positive control; HP, high positive control. ..... of circulating hepatitis B viral load. Gastroenterology 2006;130:678-686. 4.