Simultaneous detection of SENV-D and SENV-H with polymerase chain reaction

  Objective   To establish a nested polymerase chain reaction(nPCR)for simultaneous detection of SENV-D and H.

  Methods   The outer primers of the first-round PCR were derived from the conserved region of ORF1 of SENV-D and H.Two sets of specific inner primers of the second-round PCR were obtained from ORF1 of SENV-D and SENV-H, respectively.The products of the nPCR were cloned into plasmids and sequenced by the dideoxy-mediated chain-termination method.

  Results   The maximum dilution of serum was 10³ for detection of SENV-D and H using the nPCR.Sequence analysis showed the nucleotide homology was 92% between the cloned SENV strains(DTd and DTh)and SENV-D and SENV-H registered in GenBank, respectively.The total rate of SENV-D and H infections was 58.4% in patients with chronic hepatitis B.

  Conclusion   This method can be used for the clinical diagnosis and epidemiological study of SENV-D and H infections.