Detection of BCR-ABL fusion gene by multiplex RT-PCR combining with capillary electrophoresis

doi:10.3969/j.issn.1673-8640.2014.03.006

Abstract

Abstract:

Objective　To establish a method for detecting BCR-ABL fusion gene by multiplex reverse transcription polymerase chain reaction ( RT-PCR),and to provide a useful tool for chronic myeloid leukemia (CML) auxiliary diagnosis. Methods　The construction of BCR-ABL fusion gene as positive template was made by overlap extension method. The method of multiplex RT-PCR combining with capillary electrophoresis was designed and optimized. Its primary performance was evaluated. Results　The lower detection limit for BCR-ABL fusion gene (ela2,e13a2 and e14a2) by multiplex RT-PCR was 10²-10³ copies/μL. The positive rate of this method to detect 50 CML patients was 86.0%(e13a2: 20.0% and e14a2: 66.0%). Compared with the results of chromosome karyotype analysis,the positive coincidence rate was 97.6%,and the negative coincidence rate was 75.0%. The 2 methods have high consistency (Kappa=0.765,P>0.05). Conclusions　The method of multiplex RT-PCR combining with capillary electrophoresis to detect BCR-ABL fusion gene is simple and rapid with good specificity and sensitivity and is very suitable for the clinical detection of CML.

Key words: BCR-ABL fusion gene, Chronic myeloid leukemia, Multiplex reverse transcription polymerase chain reaction, Capillary electrophoresis

CLC Number:

Q503

Lü Xiaonan，YE Huiming，GU Long，ZENG Jimeng. Detection of BCR-ABL fusion gene by multiplex RT-PCR combining with capillary electrophoresis[J]. , 2014, 29(3): 226-232.

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