Molecular Detection of Drug Resistant Mutants<<Return to Laboratory
Molecular detection of nucleoside/nucleotide analog resistance by line probe hybridization assay or by sequence analysis.
- Hepatitis B
Serum or plasma sample. Minimum volume required for serum or plasma – 1.0 mL.
Collect blood in serum separator tubes (SST) or EDTA tubes.
Store samples frozen until shipped for testing. Ship frozen on dry ice.
Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.
For additional guidance on the transport of infectious substances in other languages, please click on the link below.
Patient on previous or current antiviral therapy for Hepatitis B infection. For patients on current treatment, HBV DNA viral load levels suggest antiviral failure (breakthrough) or nonresponse (i.e., confirmed rise on 2 occasions at least 1 month apart in HBV DNA by > 1 log10 IU/ml from treatment initiation, or HBV DNA reduction < 1 log10 IU/ml, respectively). Previous antiviral treatment warrants drug resistance testing to determine subsequent therapies. HBV DNA results must be positive (i.e, must have detectable viral load).
Completed Viral Hepatitis and Bloodborne Pathogens requisition including sender name, address and telephone number. Patient name or identifier (referring specimen lab #), date of birth, suspected exposure, test(s) requested. Type of specimen and date collected. If possible, include the clinical history and lab results that have already been done at local or provincial laboratories.
Conventional PCR followed by line probe hybridization assay or by sequence alignment and analysis in comparison to HBV reference sequences. Reactivity to drug resistance or compensatory mutation specific probes indicates resistance or intermediate sensitivity to certain antiviral therapies dependent on the mutation site (Lamivudine resistance: rtL80V/I, rtV/G173L, rtL180M, rtA181T/V, rtM204V/I/S; Adefovir resistance: rtA181T/V, rtN236T; Entecavir resistance: rtL180M, rtT184S/C/G/A/I/L/F/M, rtS202G/C/I, rtM204I/V, rtM250V/I/L; Tenofovir resistance or reduced susceptibility: rtA181T/V, rtA194T, rtN236T.
19 calendar days.
- Mese S, Arikan M, Cakiris A, et al. Role of the line probe assay INNO-LiPA HBV DR and ultradeep pyrosequencing in detecting resistance mutations to nucleoside/nucleotide analogues in viral samples isolated from chronic hepatitis B patients. J Gen Virol 2013 94:2729-2738.
- Degertekin B, Hussain M, Tan J, et al. Sensitivity and accuracy of an updated line probe assay (HBV DR v.3) in detecting mutations associated with hepatitis B antiviral resistance. J Hepatol 2009. 50:42-48.