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.
OBJECTIVE
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.


CONCLUSIONS
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.
Yang
Jie-Zuan
JZ
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Department of Infectious Diseases, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou.
Li
Ming-Wei
MW
Wang
Jian-Guo
JG
Lu
Hai-Feng
HF
Yao
Xin-Sheng
XS
He
Jian-Qin
JQ
Li
Lan-Juan
LJ