Lab Med Online.  2011 Oct;1(4):195-201. 10.3343/lmo.2011.1.4.5.

Analysis of Immunoglobulin Gene Rearrangement: Comparison between BIOMED-2 Multiplex PCR and Conventional Nested PCR

• Sung JY
• Kang SY
• Kim SH
• Kwon JE
• Ko YH

• KMID: 1993637
• DOI: http://doi.org/10.3343/lmo.2011.1.4.5

Abstract

BACKGROUND
Immunoglobulin (Ig) gene rearrangement analysis is a useful additional tool to detect clonality of B-lymphoproliferative disease and the method to detect immunoglobulin gene rearrangement is required the high sensitivity and specificity. BIOMED-2 multiplex PCR was designed for the evaluation of molecular clonality of lymphoid lesions. We evaluated the usefulness of the BIOMED-2 multiplex PCR by comparing it with conventional nested PCR.
METHODS
Sixteen patients with malignant lymphoma and 5 with reactive lymph nodes were examined to assess the sensitivity, specificity, and accuracy between conventional nested PCR and BIOMED-2. All 3 tests performed using the BIOMED-2 kit for immunoglobulin (Ig) heavy chain (IGH) gene, Igkappa light chain (IGK) gene, and Iglambda light chain (IGL) gene, were used to analyze clonality.
RESULTS
Both the methods showed 100% specificity (95% confidence interval, 56.6-100.0). The combination of IGH and IGK BIOMED-2 tests with or without IGL revealed the highest sensitivity (87.5%; range, 64.0-96.5%) and accuracy (90%; range, 0.70-0.97). Compared to the conventional method, the BIOMED-2 test for IGH showed a higher sensitivity (62.5%; range, 38.6-81.5%) and accuracy (71%, 0.50-0.86).
CONCLUSIONS
These results suggest that, compared to the conventional method, BIOMED-2 has higher sensitivity and allows for easier interpretation while evaluating the clonality of B-lymphoproliferative disease.

Keyword

Gene Rearrangement; Genes; Immunoglobulin; Hematologic Neoplasms; BIOMED-2

MeSH Terms

Gene Rearrangement
Genes, Immunoglobulin
Hematologic Neoplasms
Humans
Immunoglobulins
Light
Lymph Nodes
Lymphoma
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Sensitivity and Specificity
Immunoglobulins

Figure

• 

  Fig. 1 Results of both the methods in a single case. The case (case no. 9) with nodal marginal zone lymphoma (A) showed an uncertain band in the conventional method (B), but a definite peak in BIOMED-2 (C). (A, hematoxylin and eosin [H&E] stain ×40; Inset of A, H&E stain ×400; Arrow indicates uncertain band).

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