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

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Abstract
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.

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