Detection of HBsAg mutants.
HBsAg screening is carried out routinely to detect hepatitis B virus (HBV) infection. The immunoassays used employ capture antibodies often having specificity for epitopes present on the antigenic (a) determinant of the HBsAg. Loss of detection may occur due to mutations within and/or outside of the a determinant that affect conformational epitope recognition or HBsAg secretion or expression. Most of the mutations associated with immune escape occur within the second loop of the a determinant. In order to detect these HBsAg mutants, antibodies to subdominant regions within the a determinant or outside of the HBsAg may be required, and this has been the focus of many recent studies. Any changes to immunoassay formulations should also address the possible effect of HBV genotypic polymorphisms on assay specificity and sensitivity. HBsAg mutants may also be identified through nucleic acid detection of HBV in serum. Various molecular analysis methods have been developed to provide specific and sensitive detection of HBsAg mutants, including sequencing, limiting dilution cloning PCR (LDC-PCR), gap ligase chain reaction (gLCR), and real time PCR. Sequencing the HBsAg coding region provides specific information on the nucleotide sequence; however, it is relatively insensitive for the detection of minority quasispecies. Other nucleic acid methods offer greater sensitivity for the detection of point mutations. To improve immunoassays, further research will be required to increase detection sensitivity and specificity. Ultimately, a better understanding of the structure of antibody-bound HBsAg will help identify the immunological targets required for the accurate detection of HBsAg in blood.
Osiowy
Carla
C
Bloodborne Pathogens and Hepatitis, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada. carla_osiowy@phac-aspc.gc.ca
D000940:Antigenic Variation; D004279:DNA, Viral / Q000097:blood / Q000235:genetics; D005814:Genes, Viral; D006509:Hepatitis B / Q000097:blood* / Q000175:diagnosis; D006510:Hepatitis B Antibodies / Q000097:blood; D006514:Hepatitis B Surface Antigens / Q000097:blood* / Q000235:genetics / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D014766:Viremia / Q000097:blood / Q000175:diagnosis*
D004279:DNA, Viral; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens