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Hepatitis B (HBV) DNA & Mutation Analysis Service

Contract Research Services Gene Testing Services Mutation Detection Services HBV DNA & Mutation Analysis Plant Gene Analysis Custom Mutation Assays Gene Cloning & Mutagenesis Nucleic Acid Purification

Direct quantification of mutant HBV DNAs

Mutant forms of HBV are involved in drug resistance and disease progression.  For example, the rt204 mutations in the viral polymerase YMDD encoding region is involved in resistance to lamivudine, telbivudine and entecavir, while the basal core promoter (BCP) A1762T/G1764A mutation and the precore G1896A mutation are potential markers for HBeAg seroconversion [1].  Quantitative detection of these mutations can facilitate early detection of drug resistance and determination of when to initiate treatment.  However, HBV genome is notorious for being polymorphic; this poses tremendous challange to measure mutant viruses as the unpredictable mutations around the target mutation site will affect assay accuracy.  Recent advances in detection methodology allowed reliable quantification of HBV mutants [2, 3].  Further more, these ultrasensitivie technology allows early detection and monitoring of mutant viruses.

We offer ultrasensitive (0.01-0.001% mutant) and quantitative analysis of HBV mutations.  The target mutations include the rt204(ATT), rt204(ATA), rt204(ATC), rt204(GTG), A1762T/G1764A, G1896A, and G1896A/G1899A mutations.

We also offer a testing service using a set of highly sensitive (1%) quantitative assays for L180M, A181T, A181V, M204V(GTG), M204I(ATT, ATA, ATC), and N236T mutations that are directly involved in the resistance to lamivudine, telbivudine, entecavir and adefovir.  These assays offer the advantage of closed tube amplification and hybridization at accurately controlled temperatures for more accurate mutation analysis than the colometric hybridization tests.

HBV total viral load analysis

We offer a total HBV DNA (viral load) analysis service.  This test is uniquely designed to maximize the chance of viral DNA amplification and thus minimizes the fasle negatives.  Change of HBV total viral load is an important measure of treatment efficacy.  An increase of total viral load duirng treatment is a non-specific, and often late, indicator of drug resistance.

 

References:

1. Nie, H., et al: Quantitative dynamics of hepatitis B basal core promoter and precore mutants before and after HBeAg seroconversion.  Journal of Hepatology. 56:795-802, 2012.

2. Nie, H., et al: Quantification of complex precore mutations of hepatitis B virus by SimpleProbe real time PCR and dual melting analysis.  Journal of Clinical Virology. 51(4):234-240, 2011

3. Nie, H.,et al: Ultrasensitive quantification of hepatitis B virus A1762T/G1764A mutant by a SimpleProbe PCR using a wild-type-selective PCR blocker and a primer-blocker-probe partial-overlap approach.  Journal of Clinical Microbiology. 49(7):2440-2448, 2011

 


   
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