A simple method for the detection of neurologic disorders associated with CAG repeat expansion using PCR-microtiter plate hybridization

• Published in: Journal of biotechnology Vol. 95; no. 3; pp. 215 - 223
• Main Authors: Lee, Y., Oh, M.R., Kim, C.H., Hwang, H.Z., Kim, J.S., Song, S.M., Jin, D.K.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 23.05.2002; Amsterdam Elsevier; New York, NY
• Subjects: Alleles; Ataxin-3; Biological and medical sciences; Biotechnology; CAG repeat related disorders; Deoxyribonucleases, Type II Site-Specific - metabolism; DNA Probes; Fundamental and applied biological sciences. Psychology; Genetic technics applied to diagnosis; Genetic Testing; Health. Pharmaceutical industry; Humans; Huntington disease; Huntington Disease - diagnosis; Huntington Disease - genetics; Industrial applications and implications. Economical aspects; Machado-Joseph Disease - diagnosis; Machado-Joseph Disease - genetics; Microtiter plate hybridization; Nerve Tissue Proteins - genetics; Nuclear Proteins; Polymerase Chain Reaction; Repeat expansion detection; Repressor Proteins; Sensitivity and Specificity; Spinocerebellar ataxia; Spinocerebellar Degenerations - diagnosis; Spinocerebellar Degenerations - genetics; Trinucleotide Repeats; CAG repeat related disorders; Microtiter plate hybridization; Spinocerebellar ataxia; Repeat expansion detection; Huntington disease; Polymerase chain reaction; Molecular hybridization; Nervous system diseases; Microtiter plate; Microsatellite DNA; Degenerative disease; Diagnosis; Method; Detection; Expansion
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/S0168-1656(02)00024-X

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.