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Year : 2009  |  Volume : 27  |  Issue : 2  |  Page : 111--115

Quantitation of hepatitis B virus DNA in plasma using a sensitive cost-effective DQin-houseDQ real-time PCR assay

Hubert Darius J Daniel1, John G Fletcher1, George M Chandy2, Priya Abraham1 
1 Department of Clinical Virology, Christian Medical College, Vellore - 632 004, India
2 Department of Clinical Gastroenterology, Christian Medical College, Vellore - 632 004, India

Correspondence Address:
Priya Abraham
Department of Clinical Virology, Christian Medical College, Vellore - 632 004
India

Background: Sensitive nucleic acid testing for the detection and accurate quantitation of hepatitis B virus (HBV) is necessary to reduce transmission through blood and blood products and for monitoring patients on antiviral therapy. The aim of this study is to standardize an DQin-houseDQ real-time HBV polymerase chain reaction (PCR) for accurate quantitation and screening of HBV. Materials and Methods: The DQin-houseDQ real-time assay was compared with a commercial assay using 30 chronically infected individuals and 70 blood donors who are negative for hepatitis B surface antigen, hepatitis C virus (HCV) antibody and human immunodeficiency virus (HIV) antibody. Further, 30 HBV-genotyped samples were tested to evaluate the DQin-houseDQ assaySQs capacity to detect genotypes prevalent among individuals attending this tertiary care hospital. Results: The lower limit of detection of this DQin-houseDQ HBV real-time PCR was assessed against the WHO international standard and found to be 50 IU/mL. The interassay and intra-assay coefficient of variation (CV) of this DQin-houseDQ assay ranged from 1.4% to 9.4% and 0.0% to 2.3%, respectively. Virus loads as estimated with this DQin-houseDQ HBV real-time assay correlated well with the commercial artus HBV RG PCR assay ( r = 0.95, P < 0.0001). Conclusion: This assay can be used for the detection and accurate quantitation of HBV viral loads in plasma samples. This assay can be employed for the screening of blood donations and can potentially be adapted to a multiplex format for simultaneous detection of HBV, HIV and HCV to reduce the cost of testing in blood banks.

How to cite this article:
Daniel HJ, Fletcher JG, Chandy GM, Abraham P. Quantitation of hepatitis B virus DNA in plasma using a sensitive cost-effective "in-house" real-time PCR assay.Indian J Med Microbiol 2009;27:111-115

How to cite this URL:
Daniel HJ, Fletcher JG, Chandy GM, Abraham P. Quantitation of hepatitis B virus DNA in plasma using a sensitive cost-effective "in-house" real-time PCR assay. Indian J Med Microbiol [serial online] 2009 [cited 2020 Feb 25 ];27:111-115
Available from: http://www.ijmm.org/article.asp?issn=0255-0857;year=2009;volume=27;issue=2;spage=111;epage=115;aulast=Daniel;type=0