A simple method for the detection of neurologic disorders associated with CAG repeat expansion using PCR-microtiter plate hybridization.
A new screening method was developed for the detection of CAG expanded alleles in patients with hereditary ataxia using polymerase chain reaction-based microtiter plate hybridization (PCR-MPH). The system can be applied to detect pathologic alleles by hybridization with the immobilized (CAG)48 repeat probe derived from the unrelated gene 'ERDA1' except for the CAG repeats. We examined 10 individuals with SCA3, 10 with Huntington disease and 30 normal controls (31 controls for SCA3) using this method. The results showed that a clear discrimination was possible in all cases. We suggest that this system be made available for mass screening of patients with hereditary ataxia disorders. This report is the first to demonstrate that a PCR-MPH system can be successfully applied to DNA size differentiation in addition to base pair mismatches. Also, our design of the probe is unique in that the probe motif stem from the unrelated gene sequence and not from the synthetic oligonucleotides.
Lee
Y
Y
Department of Clinical Research Center, Samsung Biomedical Research Institute, Sungkyunkwan University, Samsung Medical Center, 50 Ilwon-dong, kangnam-ku, 135-710, Seoul, South Korea.
Oh
M R
MR
Kim
C H
CH
Hwang
H Z
HZ
Kim
J S
JS
Song
S M
SM
Jin
D K
DK
D000483:Alleles; D000067699:Ataxin-3; D015342:DNA Probes; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005820:Genetic Testing; D006801:Humans; D006816:Huntington Disease / Q000175:diagnosis / Q000235:genetics; D017827:Machado-Joseph Disease / Q000175:diagnosis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics; D009687:Nuclear Proteins; D016133:Polymerase Chain Reaction; D012097:Repressor Proteins; D012680:Sensitivity and Specificity; D013132:Spinocerebellar Degenerations / Q000175:diagnosis* / Q000235:genetics; D018911:Trinucleotide Repeats
D015342:DNA Probes; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; C041749:endodeoxyribonuclease SfiI; D015252:Deoxyribonucleases, Type II Site-Specific; C092341:ATXN3 protein, human; D000067699:Ataxin-3