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Comparison of the sensitivity of NAT using pooled donor samples for HBV and that of a serologic HBsAg assay
Author(s) -
Sato Shinichiro,
Ohhashi Wataru,
Ihara Hiromi,
Sakaya Shinichi,
Kato Toshiaki,
Ikeda Hisami
Publication year - 2001
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.2001.41091107.x
Subject(s) - serial dilution , hbsag , seroconversion , serology , virology , medicine , hepatitis b virus , immunoassay , primer (cosmetics) , antibody , immunology , chemistry , virus , pathology , alternative medicine , organic chemistry
BACKGROUND : Studies were conducted using samples from early and late‐stage HBV‐infected persons to determine the pool size at which PCR had better sensitivity than a sensitive HBsAg chemoluminescence immunoassay (CLIA‐HBsAg). STUDY DESIGN AND METHODS: HBV seroconversion panels were tested for HBsAg by CLIA and for HBV DNA by nested PCR (95% hit rate: 100 copies/mL); PCR was carried out at various dilutions. HBV serologically positive samples that were detected from the simultaneous screening of 540,161 routine whole‐blood donations using CLIA‐HBsAg and agglutination assays were also characterized for additional markers of HBV infection. RESULTS: In 9 of 10 HBV seroconversion panels, PCR had better sensitivity than CLIA‐HBsAg at dilutions of 1‐in‐25 or lower. Of 65 CLIA‐only confirmed‐positive donor samples (agglutination assay‐negative), 8 represented early infection, 2 of which were PCR positive at a 1‐in‐50 dilution but negative at a 1‐in‐100 dilution. Only 2 of 47 samples from probable late‐stage HBV infection that were positive on CLIA only were PCR positive with 0.1‐mL sample volume and the S‐region primer; the remaining 45 samples required a 1.0‐mL sample input and C‐region primer for increased PCR positivity. The remaining 10 CLIA‐only confirmed‐positive donor samples were from HBV vaccine recipients. None of the 12 CLIA‐ and HBsAg‐negative donor samples that were strongly anti‐HBc reactive could be detected by PCR at any dilution; all 12 were PCR positive when undiluted, but 4 required a 1.0‐mL input volume for PCR positivity. CONCLUSION: For the detection of samples representing early‐stage HBV infection, PCR at dilutions of 1‐in‐25 or lower (equivalent to a pool of ≤25 members) had greater sensitivity than CLIA‐HBsAg. In contrast, samples from late‐stage HBV infection were detected by PCR only with undiluted samples (0.1‐mL or 1.0‐mL input volumes), regardless of CLIA‐HBsAg reactivity. Therefore, although NAT using minipools of 25 donations or less may be effective for the detection of early‐stage HBV infection, it may not be effective for the detection of persistent HBV infection.