ARMS-PCR combined with capillary electrophoresis can be a sensitive and quantitative method for detection of MYD88-L265P mutation in lymphoma.
10.7534/j.issn.1009-2137.2018.06.014
- Author:
Zi-Xuan DING
1
;
Hong LIU
1
;
Jun-Dan XIE
1
;
Hong YAO
1
;
Liang MA
1
;
Qiao-Chen QIU
1
;
Hong-Jie SHEN
2
Author Information
1. The First Affiliated Hospital of Soochow University, Suzhou 215006,China；Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Collaborative Innovation Center of Hematology, Suzhou 215006, China.
2. The First Affiliated Hospital of Soochow University, Suzhou 215006,China；Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Collaborative Innovation Center of Hematology, Suzhou 215006, China.E-mail:shj98538@sina.com.
- Publication Type:Journal Article
- MeSH:
DNA Mutational Analysis;
Electrophoresis, Capillary;
Humans;
Lymphoma;
Mutation;
Myeloid Differentiation Factor 88;
genetics;
Polymerase Chain Reaction
- From:
Journal of Experimental Hematology
2018;26(6):1663-1667
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the feasibility of sensitive and quantitative detection of MYD88 gene L265P mutation in lymphoma patients by using ARMS-PCR combined with capillary electrophoresis.
METHODS:ARMS-PCR amplified MYD88 gene was analyzed by capillary electrophoresis in ABI 3730 sequencer; Exon 5 of the same gene was sequenced bi-directionally as reported.
RESULTS:The sensitivity of detection L265P mutations by the ARMS-PCR combined with capillary electrophoresis and direct sequencing was 0.2% and 5%, respectively, according to the detection of the gradient-diluted plasmid standards. The detection rate of 184 patients was 13.59% and 8.28%, respectively (p<0.001). Moreover, the former method can successfully detect the mutation ratio(R=0.979), and the repeatabilities (CV=2.86%, 1.94%, 5.49%) are acceptable.
CONCLUSION:ARMS-PCR combined with capillary electrophoresis can quantitatively detect the MYD88 gene L265P mutation, and the detection sensitivity is significantly higher than sanger sequencing. As a supplement to the latter, it can effectively lead to the earlier diagnose and monitoring of minimal residual disease.
