Premium
Rapid detection of clonal expansion of T‐cell receptor‐beta gene in patients with HBV using the real‐time PCR with DNA melting curve analysis
Author(s) -
Yang JieZuan,
Li MingWei,
Wang JianGuo,
Lu HaiFeng,
Yao XinSheng,
He JianQin,
Li LanJuan
Publication year - 2010
Publication title -
hepatology research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.123
H-Index - 75
eISSN - 1872-034X
pISSN - 1386-6346
DOI - 10.1111/j.1872-034x.2009.00600.x
Subject(s) - gene , biology , microbiology and biotechnology , polymerase chain reaction , t cell receptor , dna , real time polymerase chain reaction , t cell , genetics , immune system
Aim: The gene melting spectral pattern (GMSP) of PCR products from 24 T‐cell receptor beta chain variable ( TCRBV ) gene families was developed to determine sequence bias and feature of TCRBV CDR3 gene family. Methods: The assay was based on reverse transcript quantitative polymerase chain reaction and their DNA melting curves. Results: We discovered that the relatively conserved amino acid sequences X‐Q and X‐G are present in TCRBV CDR3 from patients with HBV. Further, the X of the X‐Q motif is preferentially E (glutamic acid), P (proline) or T (threonine) when accompanied by the BJ2.7, BJ1.5, or BJ2.3, respectively. The frequency of sequence bias in the TCRBV gene family showed a positive correlation with the T cell receptor excision circles (TRECs) content, and an inverse correlation with the HBV DNA loading. Conclusion: These results suggest that the GMSP assay could be used to monitor the features of TCRBV gene distribution quickly, and facilitate the further study of HBV‐specific T cell in patients with HBV.