면역글로불린 유전자 재배열 분석법: BIOMED-2와 통상적 Nested PCR법의 비교
Analysis of Immunoglobulin Gene Rearrangement: Comparison between BIOMED-2 Multiplex PCR and Conventional Nested PCR
1
조회
0
다운로드
3
인용
0
참고문헌
초록
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, Igκ light chain (IGK) gene, and Igλ 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.
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, Igκ light chain (IGK) gene, and Igλ 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.
참고문헌 (21)
- Roman E, "Epidemiology of lymphomas", Histopathology, 58, pp.4~14 (2011)
- Cook JR, "Cell and tissue based molecular pathology: a volume in the series foundations in diagnostic pathology", pp.305~324 (2008), Churchill Livingstone
- Davis TH, "Detection of clonal immunoglobulin gene rearrangements by polymerase chain reaction amplification and single-strand conformational polymorphism analysis", Am J Pathol, 142, pp.1841~1847 (1993)
- Linke B, "Automated high resolution PCR fragment analysis for identification of clonally rearranged immunoglobulin heavy chain genes", Leukemia, 11, pp.1055~1062 (1997)
- van Dongen JJ, "Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED- 2 concerted action BMH4-CT98-3936", Leukemia, 17, pp.2257~2317 (2003)
- Burack WR, "PCR assays detect B-lymphocyte clonality in formalin-fixed, paraffin-embedded specimens of classical hodgkin lymphoma without microdissection", Am J Clin Pathol, 134, pp.104~111 (2010)
- Evans PA, "Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED- 2 concerted action BHM4-CT98-3936", Leukemia, 21, pp.207~214 (2007)
- Hebeda KM, "PCR clonality detection in Hodgkin lymphoma", J Hematop, 2, pp.34~41 (2009)
- 오혜룡, "카파 및 람다 면역글로불린 경쇄 유전자의 클론성 재배열을 동시에 보이는 람다 경쇄 표현 폐 MALT 림프종 1예", Annals of Laboratory Medicine, 29(3), pp.256~261 (2009), 대한진단검사의학회
- Achille A, "Routine application of polymerase chain reaction in the diagnosis of monoclonality of B-cell lymphoid proliferations", Diagn Mol Pathol, 4, pp.14~24 (1995)
- Chute DJ, "Detection of immunoglobulin heavy chain gene rearrangements in classic hodgkin lymphoma using commercially available BIOMED-2 primers", Diagn Mol Pathol, 17, pp.65~72 (2008)
- Male D, "Immunology", pp.80~85 (2006), Mosby
- Bagg A, "Immunoglobulin heavy chain gene analysis in lymphomas: a multi-center study demonstrating the heterogeneity of performance of polymerase chain reaction assays", J Mol Diagn, 4, pp.81~89 (2002)
- Tümkaya T, "Identification of immunoglobulin lambda isotype gene rearrangements by Southern blot analysis", Leukemia, 10, pp.1834~1839 (1996)
- van der Burg M, "Ordered recombination of immunoglobulin light chain genes occurs at the IGK locus but seems less strict at the IGL locus", Blood, 97, pp.1001~1008 (2001)
- van Krieken JH, "Improved reliability of lymphoma diagnostics via PCR-based clonality testing: report of the BIOMED-2 concerted action BHM4-CT 98-3936", Leukemia, 21, pp.201~206 (2007)
- Halldórsdóttir AM, "Application of BIOMED-2 clonality assays to formalin-fixed paraffin embedded follicular lymphoma specimens: superior performance of the IGK assays compared to IGH for suboptimal specimens", Leuk Lymphoma, 48, pp.1338~1343 (2007)
- Smilevska T, "Immunoglobulin kappa gene repertoire and somatic hypermutation patterns in follicular lymphoma", Blood Cells Mol Dis, 41, pp.215~218 (2008)
- Liu H, "A practical strategy for the routine use of BIOMED-2 PCR assays for detection of B- and T-cell clonality in diagnostic haematopathology", Br J Haematol, 138, pp.31~43 (2007)
- Kros JM, "Analysis of immunoglobulin H gene rearrangement by polymerase chain reaction in primary central nervous system lymphoma", J Neurosurg, 97, pp.1390~1396 (2002)
- Essop MF, "Analysis of low and high grade B-cell lymphoma subtypes using semi-nested PCR and two primer sets", Eur J Haematol, 59, pp.136~141 (1997)