[Primers for detecting gene rearrangement in different regions of immunoglobulin heavy chain genes and their application in diagnosis of paraffin-embedded lymphoma tissues].
OBJECTIVE
To analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.


METHODS
Three pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.


RESULTS
The positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.


CONCLUSIONS
Primers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.
Qi
Zong-Li
ZL
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. qizongli@126.com
Zhang
Bao
B
Han
Xi-Qun
XQ
Zhu
Mei-Gang
MG
Zhao
Tong
T
D017931:DNA Primers; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D016393:Lymphoma, B-Cell / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D008228:Lymphoma, Non-Hodgkin / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D016399:Lymphoma, T-Cell / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D008297:Male; D016612:Paraffin Embedding
D017931:DNA Primers; D007143:Immunoglobulin Heavy Chains