[Detection of minimal residual disease in acute lymphoblastic leukemia using PCR-molecule hybridization of RNA transcripts].
OBJECTIVE
To detect acute lymphoblastic leukemia's(ALL's) minimal residual disease(MRD) rapidly and effectively.


METHODS
In this assay, the gamma T-cell receptor gene rearrangements serve as marker genes. The gene rearrangements are amplified from the diagnostic specimens using a consensus V segment primer and a consensus J segment primer to which the promoter T7 RNA polymerase has been appended. The PCR product from this amplification is transcribed into a radiolabeled RNA probe. The opposite DNA strand is transcribed into test RNA from the PCR product of different staged specimens. The test RNA is hybridized with the probe, and later the digestion with RNase A, Polyacrylamide gel electrophoresis and autoradiography are in progress.


RESULTS
According to the mechanism, the perfectly matched RNA duplex can prevent the digestion of RNase A, and the presence of the leukemia cells in the test specimen can be determined. Logarithmical dilution experiments with DNA of a cell line from ALL have shown that this assay's sensitivity is at the 10(-5) level. Minimal residual disease was successfully detected in a case of ALL during its complete remission stage. But if the probe and test RNA are not from the same individual, the results of this kind of cross hybridization are negative.


CONCLUSIONS
The above results suggest that this assay can become an effective measure in the detection of ALL-MRD clinically.
Liang
X
X
Department of Pediatrics, Union Hospital, Tongji Medical University, Wuhan, Hubei Province, 430022 P.R.China.
Fei
H
H
Xiao
Y
Y
Liu
S
S
Yang
A
A
D002648:Child; D005260:Female; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D006801:Humans; D018365:Neoplasm, Residual; D016133:Polymerase Chain Reaction; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis*; D012333:RNA, Messenger / Q000235:genetics*; D012680:Sensitivity and Specificity
D012333:RNA, Messenger