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The evaluation of minimal residual disease in multiple myeloma by fluorescent molecular beacons in real time PCR of IgH gene rearrangements and correlation with flow cytometry.
J BUON. 2013 Apr-Jun; 18(2):442-7.JB

Abstract

PURPOSE

Multiple myeloma (MM) patients relapse after a period of time despite longer disease-free survival due to novel treatment options. In this study we aimed to assess the value of real-time polymerase chain reaction (RT-PCR) for detecting the immunoglobulin heavy chain (IgH) gene rearrangement using allele-specific molecular beacons as fluorescence probes to quantify minimal residual disease (MRD) and also to correlate post-treatment flow cytometric detection of plasma cells' (PCs) expression of CD19, CD38, CD45, CD56 and CD138 in MM.

METHODS

After diagnosis of 17 MM patients, the CDR1, CDR2 and CDR3 regions of the IgH gene were analysed and sequenced to identify IgH's clonal nature. Unique sequences of the clonal IgH rearrangement were used to design specific molecular beacon probes for each MM patient. Examined were also the co-expression of CD19, CD38, CD45, CD56, and CD138 molecules in bone marrow aspirates of patients with MM by flow cytometry.

RESULTS

Detection of MRD was positive in 13 (76%) of 17 patients by RT-PCR. The infiltration ratio was significantly correlated with CD138 expression (p=0.009). Significant correlation was also found between RT-PCR detection of MRD and CD138 expression (p=0.006). Nevertheless, no correlation was observed among other surface antigens (CD38, CD45, CD56).

CONCLUSION

Our results indicated that RT-PCR with specific molecular beacons provide a feasible, accurate and reproducible method for the determination of MRD in MM. Flow cytometry detection of CD138 expression may be used as a disease marker in addition to RT-PCR.

Authors+Show Affiliations

Cukurova University Medical Faculty, Department of Medical Oncology, Adana, Turkey.No affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

23818359

Citation

Kara, I O., et al. "The Evaluation of Minimal Residual Disease in Multiple Myeloma By Fluorescent Molecular Beacons in Real Time PCR of IgH Gene Rearrangements and Correlation With Flow Cytometry." Journal of B.U.ON. : Official Journal of the Balkan Union of Oncology, vol. 18, no. 2, 2013, pp. 442-7.
Kara IO, Duman BB, Afsar CU. The evaluation of minimal residual disease in multiple myeloma by fluorescent molecular beacons in real time PCR of IgH gene rearrangements and correlation with flow cytometry. J BUON. 2013;18(2):442-7.
Kara, I. O., Duman, B. B., & Afsar, C. U. (2013). The evaluation of minimal residual disease in multiple myeloma by fluorescent molecular beacons in real time PCR of IgH gene rearrangements and correlation with flow cytometry. Journal of B.U.ON. : Official Journal of the Balkan Union of Oncology, 18(2), 442-7.
Kara IO, Duman BB, Afsar CU. The Evaluation of Minimal Residual Disease in Multiple Myeloma By Fluorescent Molecular Beacons in Real Time PCR of IgH Gene Rearrangements and Correlation With Flow Cytometry. J BUON. 2013 Apr-Jun;18(2):442-7. PubMed PMID: 23818359.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The evaluation of minimal residual disease in multiple myeloma by fluorescent molecular beacons in real time PCR of IgH gene rearrangements and correlation with flow cytometry. AU - Kara,I O, AU - Duman,B B, AU - Afsar,C U, PY - 2013/7/3/entrez PY - 2013/7/3/pubmed PY - 2013/11/20/medline SP - 442 EP - 7 JF - Journal of B.U.ON. : official journal of the Balkan Union of Oncology JO - J BUON VL - 18 IS - 2 N2 - PURPOSE: Multiple myeloma (MM) patients relapse after a period of time despite longer disease-free survival due to novel treatment options. In this study we aimed to assess the value of real-time polymerase chain reaction (RT-PCR) for detecting the immunoglobulin heavy chain (IgH) gene rearrangement using allele-specific molecular beacons as fluorescence probes to quantify minimal residual disease (MRD) and also to correlate post-treatment flow cytometric detection of plasma cells' (PCs) expression of CD19, CD38, CD45, CD56 and CD138 in MM. METHODS: After diagnosis of 17 MM patients, the CDR1, CDR2 and CDR3 regions of the IgH gene were analysed and sequenced to identify IgH's clonal nature. Unique sequences of the clonal IgH rearrangement were used to design specific molecular beacon probes for each MM patient. Examined were also the co-expression of CD19, CD38, CD45, CD56, and CD138 molecules in bone marrow aspirates of patients with MM by flow cytometry. RESULTS: Detection of MRD was positive in 13 (76%) of 17 patients by RT-PCR. The infiltration ratio was significantly correlated with CD138 expression (p=0.009). Significant correlation was also found between RT-PCR detection of MRD and CD138 expression (p=0.006). Nevertheless, no correlation was observed among other surface antigens (CD38, CD45, CD56). CONCLUSION: Our results indicated that RT-PCR with specific molecular beacons provide a feasible, accurate and reproducible method for the determination of MRD in MM. Flow cytometry detection of CD138 expression may be used as a disease marker in addition to RT-PCR. SN - 1107-0625 UR - https://www.unboundmedicine.com/medline/citation/23818359/The_evaluation_of_minimal_residual_disease_in_multiple_myeloma_by_fluorescent_molecular_beacons_in_real_time_PCR_of_IgH_gene_rearrangements_and_correlation_with_flow_cytometry_ L2 - https://www.jbuon.com/archive/18-2-442.pdf DB - PRIME DP - Unbound Medicine ER -