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Objective To investigate the clonal rearrangement characteristics and clinical application value of IGH gene in B-cell non-Hodgkin’s lymphoma (B-NHL). Methods Demographic and clinical data as well as IGH sequencing results of 55 patients with B-NHL who underwent next-generation sequencing (NGS) testing were collected, and IGH gene clonal rearrangement was detected. The characteristics of IGH gene clonal rearrangement, IGHV gene usage, and the clinical application value of NGS for IGH clonal rearrangement were analyzed. Results Among 55 patients with B-NHL and IGH clonal rearrangement, single dominant clones were mainly detected (85.45%, 47/55); a few patients had two (12.73%, 7/55) and three dominant clones (1.82%, 1/55). In terms of preference for IGHV gene usage, IGHV3 gene had the highest frequency of access in B-NHL, followed by IGHV4. Among the IGHV subtypes, IGHV3-23 had the highest frequency in chronic lymphocytic leukemia/small lymphocytic lymphoma, and IGHV4-34 had the highest frequency in primary central nervous system diffuse large B-cell lymphoma and not otherwise specified diffuse large B-cell lymphoma. Conclusion A preference for IGHV gene usage in clonal rearrangement of IGH genes is noted in B-NHL patients with different pathological types. Using NGS to detect IGH clonal rearrangement can identify subclones and clonal correlations, and assist in disease diagnosis. [ABSTRACT FROM AUTHOR]