HBV DNA Test Among Blood Donations May Require Two Amplification Targets
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To analyze the risk and reason of false-negative HBV DNA results of NAT reagents among blood donations of China and discuss the necessity of two amplification targets for HBV DNA tests among donations. In this study, samples that showed discordant results on two commercially available assay platforms were further detected by established in-house methods based on conserved regions of the HBV genome. The HBV concentration of these samples was determined using two commercially available reagents. The samples with high titers of HBV were detected by an in-house method. The samples showing high Ct differences between two regions in the in-house method were further sequenced and aligned with primers and probes. The results showed that the established method has a good detection performance. The mismatch between reverse primers and sample sequences led to decreased detection capacity of S and C regions by the in-house method, but it could be compensated by another region. Among the false-negative samples detected by commercial reagents, most were because of low titers; however, there were 7 samples with HBV DNA concentrations much higher than the LOD of the commercial reagents, which may be due to mismatch of the primer or probe. This study highlights the potential risk of HBV false-negative detection by commercial NAT reagents. The dual-target assay may be helpful for HBV screening and reduce the risk of false-negative detection.
KeywordsHepatitis B virus Nucleotide acid test Dual-target False-negative Transmission risk
The authors thank the staff at the participating blood banks who are engaged in this study.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no competing interest.
The study was approved by the institutional review board of the National Center for Clinical Laboratories. Written informed consent was obtained from all subjects participating in this research.
- 2.Liang X, Bi S, Yang W, Wang L, Cui G, Cui F, Zhang Y, Liu J, Gong X, Chen Y, Wang F, Zheng H, Wang F, Guo J, Jia Z, Ma J, Wang H, Luo H, Li L, Jin S, Hadler SC, Wang Y (2009) Epidemiological serosurvey of hepatitis B in China—declining HBV prevalence due to hepatitis B vaccination. Vaccine 27:6550–6557CrossRefGoogle Scholar
- 7.Tipples GA, Ma MM, Fischer KP, Bain VG, Kneteman NM, Tyrrell DL (1996) Mutation in HBV RNA-dependent DNA polymerase confers resistance to lamivudine in vivo. Hepatology 24:714–717Google Scholar
- 16.Allain JP, Mihaljevic I, Gonzalez-Fraile MI, Gubbe K, Holm-Harritshøj L, Garcia JM, Brojer E, Erikstrup C, Saniewski M, Wernish L, Bianco L, Ullum H, Candotti D, Lelie N, Gerlich WH, Chudy M (2013) Infectivity of blood products from donors with occult hepatitis B virus infection. Transfusion 53:1405–1415CrossRefGoogle Scholar
- 17.Andonov A, Osiowy C, Borlang J, Swidinsky K (2016) Sequence variability of the Cobas taqman assay target region impacts accurate HBV DNA detection. Vox Sang 111:S58Google Scholar