Hepatitis B virions isolated with antibodies to the pre-S1 domain reveal occult viremia by PCR in Alaska Native HBV carriers who have seroconverted.
BACKGROUND
Occult viremia occurring before the appearance of HBsAg or after the disappearance of HBsAg is detectable by gene amplification technologies whose efficiency depends on nucleic acid preparation.


METHODS
To isolate HBV DNA from viremic plasma, immunoaffinity capture (IAC) of intact HBV with biotinylated pre-S1 antibodies coupled to streptavidin-coated magnetic beads was evaluated. IAC was compared with a silica-gel method (Qiagen [QSG]) and its two modifications wherein the samples were heated with lysis buffer at 60(o)C for 10 minutes (QSG-60) or at 58 degrees C for 60 minutes with proteinase-K (QSG-PK). Each HBV DNA sample was tested by heminested PCR amplification of the HBV gene sequences. A total of 36 coded serum samples were tested, including three HBsAg-positive controls and 33 former chronic HBV carriers who had seroconverted (developed antibody to HBsAg [anti-HBs]). Commercially available seroconversion panels (PHM 907, 911, and 922) were similarly tested for window-period viremia.


RESULTS
In the 33 former chronic HBV carriers who had seroconverted, IAC revealed HBV DNA in 17 samples, whereas it was revealed in only 11 samples by QSG-PK (p = 0.031), 10 by QSG-60 (p = 0.016), and 9 by QSG (p = 0.0078). However, HBV DNA was not amplified from the 17 samples at 1-in-10 dilutions; thus, they were considered to have low-level viremia. IAC revealed HBV DNA as early as or earlier than the other methods in PHM 907, 911, and 922 panels.


CONCLUSIONS
IAC is apparently an optimal method of sample preparation for amplification of HBV DNA in patients in the pre-HBsAg window period, and for detecting low-level viremia persistent in several individuals who were former chronic HBV carriers who had seroconverted (developed anti-HBs).
Gandhi
M J
MJ
Department of Laboratory Medicine and the Liver Center, University of California, San Francisco, California 94143, USA.
Yang
G G
GG
McMahon
B J
BJ
Vyas
G N
GN
D000413:Alaska / Q000453:epidemiology; D002353:Carrier State / Q000378:metabolism / Q000821:virology; D004279:DNA, Viral / Q000097:blood; D006509:Hepatitis B / Q000097:blood* / Q000453:epidemiology / Q000235:genetics; D006510:Hepatitis B Antibodies / Q000097:blood / Q000276:immunology; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D009780:Occult Blood; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary / Q000235:genetics*; D014766:Viremia / Q000235:genetics*
D004279:DNA, Viral; D006510:Hepatitis B Antibodies