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Quantifying perinatal transmission of Hepatitis B viral quasispecies by tag linkage deep sequencing

  • Author(s): Du, Y
  • Chi, X
  • Wang, C
  • Jiang, J
  • Kong, F
  • Yan, H
  • Wang, X
  • Li, J
  • Wu, NC
  • Dai, L
  • Zhang, TH
  • Shu, S
  • Zhou, J
  • Yoshizawa, JM
  • Li, X
  • Bhattacharya, D
  • Wu, TT
  • Niu, J
  • Sun, R
  • et al.

Despite full immunoprophylaxis, mother-to-child transmission (MTCT) of Hepatitis B Virus still occurs in approximately 2-5% of HBsAg positive mothers. Little is known about the bottleneck of HBV transmission and the evolution of viral quasispecies in the context of MTCT. Here we adopted a newly developed tag linkage deep sequencing method and analyzed the quasispecies of four MTCT pairs that broke through immunoprophylaxis. By assigning unique tags to individual viral sequences, we accurately reconstructed HBV haplotypes in a region of 836 bp, which contains the major immune epitopes and drug resistance mutations. The detection limit of minor viral haplotypes reached 0.1% for individual patient sample. Dominance of "a determinant" polymorphisms were observed in two children, which pre-existed as minor quasispecies in maternal samples. In all four pairs of MTCT samples, we consistently observed a significant overlap of viral haplotypes shared between mother and child. We also demonstrate that the data can be potentially useful to estimate the bottleneck effect during HBV MTCT, which provides information to optimize treatment for reducing the frequency of MTCT.

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