Application of Multiplex Reverse-Transcriptase Polymerase Chain Reaction for Identification of Leukemia Associated Gene Abnormalities

PUOPOSE: The best prognostic predictor for acute leukemia is known to be the finding of genetic abnormalities of leukemic cells. Methods for detecting the genetic abnormalities include chromosomal studies for karyotyping, FISH(Fluorescence in situ hybridization) and RT-PCR. However, each methods have limitations i.e. low sensitivity in karyotyping, uncertainty of molecular probe to be used in FISH or RT-PCR methods. Multiplex RT-PCR(MRT-PCR) allows simultaneous detection of 29 fusion genes, more than 80 break points and splice variant. Therefore, this method can be used for the detection of molecular abnormality in fresh unknown leukemic cases, as well as for molecular remission in follow-up cases. The aim was to demonstrate whether a MRT-PCR system might be successfully used to screen a large number of patients with acute leukemia and compare the result with that of chromosome studies. DESIGN AND METHOD: Frozen bone marrow cells from 78 patients, who were diagnosed with acute leukemia at Ajou university hospital between September 1994 and February 2004, were used for MRT-PCR. In all samples with known conventional cytogenetic results, we performed MRT-PCR and compared with conventional cytogenetic study regarding the concordance rate and analyzed discordant cases regarding their types. RESULTS: 78 samples(40 male and 38 female patients) were analyzed, and there were 59 AML patients and 19 ALL patients. We successfully obtained the mRNA from all frozen samples. In 21 cases, we identified gene abnormalities with chromosome studies and most of them (15/21) showed the same abnormalities with MRT-PCR. In 57 patients with normal karyotype in cytogenetic technique, 18 translocations of clinical significance by MRT-PCR method were identified. In 18 discordant cases, there were 4 cases with t(15;17), 4 cases with t(8;21), 3 cases with t(9;22), 2 cases with t(11;19) and 4 others [t(9;11),t(9;9),t(3;11),inv(16)] CONCLUSION: There were 73% concordance rate between cytogenetic technique and MRT-PCR. Furthermore, clinically significant translocations were detected by MRT-PCR in 18 out of 57 normal karyotype patients, indicating improved sensitivity with MRT-PCR. Further investigations are needed to ascertain the usefulness of MRT-PCR for the screening tool of leukemic gene abnormalities

Author dc:creator

• 정, 연무

Contributors dc:contributor

• 김, 효철
• 대학원 의학과
• 200324434

• 급성 백혈병
• 유전자 변이
• Multiplex RT-PCR
• Acute leukemia
• gene abnormality

Language dc:language

ko

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000000094
000000000094

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/2318