Detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma by hemi-nested PCR / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 589-592, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-255837
ABSTRACT
<p><b>OBJECTIVES</b>To explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.</p><p><b>METHODS</b>bcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.</p><p><b>RESULTS</b>bcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.</p><p><b>CONCLUSION</b>There are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Gene Rearrangement, B-Lymphocyte, Heavy Chain
/
Polymerase Chain Reaction
/
Lymphoma, Large B-Cell, Diffuse
/
Genes, bcl-2
/
Genetics
/
Methods
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Hematology
Year:
2005
Type:
Article
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