Korean J Hematol 1993; 28(1):
Published online March 31, 1993
© The Korean Society of Hematology
만성 골수성 백혈병에서 중합효소 연쇄반응(polymerase chain reaction)을 이용한 BCR-ABL 유전자 재배열의 관찰
김준식, 김정희, 어완규, 김선희, 김시영, 윤휘중, 조경삼
경희대학교 의과대학 내과학교실,
경희대학교 의과대학 임상병리학교실
Detection of BCR - ABL Gene Rearrangement by Use of the Polymerase Chain Reaction in Chronic Myeloid Leukemia
Background: The Philadelphia chromosome (Ph1) results from a reciprocal translocation involving chromosomes 9 and 22, t(9;22) (q34;q11). The
polymerase chian reaction (PCR) allows the detection of minimal amounts of nucleic sequences and has been successfully used to detect the chronic myeloid leukemia (CML) - specific bcr/ abl transcripts.
In this report, We studied the clinical, cytogenetic and molecular genetic features in
eighteen CML patients, three chronic myeloproliferative disease patients and two
myelodysplastic syndrome patients.
Methods: Total RNA was isolated from cells of peripheral blood or bone marrow aspirates. The cDNA was synthesized using Moloney murine leukemia virus (M -MLV) reverse transcriptase and amplified by nested PCR. The bar rearrangement was analysed by observation of ethidium bromide stained agarose gel after electrophoresis.
Results:1. In 1st step PCR, bcr rearrangement was detected in 8 of 18 CML patients and in 2nd step PCR, bcr rearrangement was found in all CML patients. The bcr rearrangement was not found in three chronic myeloproliferative disease and two
myelodyplastic syndrome.
2. In CML patients, the bcr rearrangement revealed two types of chimeric mRNA :one with 5'breakpoint (abl exon 2 linked to bar exon 2) was found in 39% (7/18), and
the other with 3'breakpoint (abl exon 2 linked to bcr exon 3) was found in 61% (11/18).
Among the 15 samples obtained in chronic phase, 5 patients had 5'breakpoint, and 10 patients had3'breakpoint. Among the 3 samples obtained in blast crisis, 1 patient had
5'breakpoint, and 2 patients had 3'breakpoint.
Conclusion: Nested polymerase chain reaction was highly sensitive for rapid detection of bcr-abl mRNA in CML patients.
Keywords Chronic myeloid leukemia; bcr-abl gene; polymerase chain reaction(PCR);
PDF
Standard view
Export citation
Share Article
Korean J Hematol 1993; 28(1): 71-80
Published online March 31, 1993
Copyright © The Korean Society of Hematology.
만성 골수성 백혈병에서 중합효소 연쇄반응(polymerase chain reaction)을 이용한 BCR-ABL 유전자 재배열의 관찰
김준식, 김정희, 어완규, 김선희, 김시영, 윤휘중, 조경삼
경희대학교 의과대학 내과학교실,
경희대학교 의과대학 임상병리학교실
Detection of BCR - ABL Gene Rearrangement by Use of the Polymerase Chain Reaction in Chronic Myeloid Leukemia
Joon Sik Kim, Jeong Hee Kim, Wan Gyu Uh, Sun Hee Kim, Si Young Kim, Hwi Joong Yoon, Kyung Sam Cho
Department of Internal Medicine, Clinical Pathology, Kyung Hee University School of Medicine, Seoul, Korea
Abstract
Background: The Philadelphia chromosome (Ph1) results from a reciprocal translocation involving chromosomes 9 and 22, t(9;22) (q34;q11). The
polymerase chian reaction (PCR) allows the detection of minimal amounts of nucleic sequences and has been successfully used to detect the chronic myeloid leukemia (CML) - specific bcr/ abl transcripts.
In this report, We studied the clinical, cytogenetic and molecular genetic features in
eighteen CML patients, three chronic myeloproliferative disease patients and two
myelodysplastic syndrome patients.
Methods: Total RNA was isolated from cells of peripheral blood or bone marrow aspirates. The cDNA was synthesized using Moloney murine leukemia virus (M -MLV) reverse transcriptase and amplified by nested PCR. The bar rearrangement was analysed by observation of ethidium bromide stained agarose gel after electrophoresis.
Results:1. In 1st step PCR, bcr rearrangement was detected in 8 of 18 CML patients and in 2nd step PCR, bcr rearrangement was found in all CML patients. The bcr rearrangement was not found in three chronic myeloproliferative disease and two
myelodyplastic syndrome.
2. In CML patients, the bcr rearrangement revealed two types of chimeric mRNA :one with 5'breakpoint (abl exon 2 linked to bar exon 2) was found in 39% (7/18), and
the other with 3'breakpoint (abl exon 2 linked to bcr exon 3) was found in 61% (11/18).
Among the 15 samples obtained in chronic phase, 5 patients had 5'breakpoint, and 10 patients had3'breakpoint. Among the 3 samples obtained in blast crisis, 1 patient had
5'breakpoint, and 2 patients had 3'breakpoint.
Conclusion: Nested polymerase chain reaction was highly sensitive for rapid detection of bcr-abl mRNA in CML patients.
Keywords: Chronic myeloid leukemia, bcr-abl gene, polymerase chain reaction(PCR),
Previous Article
