Rapid diagnosis of CMT1A duplications and HNPP deletions by multiplex microsatellite PCR.
Charcot-Marie-Tooth (CMT) disease and hereditary neuropathy with liability to pressure palsies (HNPP) are frequent forms of genetically heterogeneous peripheral neuropathies. Reciprocal unequal crossover between flanking CMT1A-REPs on chromosome 17p11.2-p12 is a major cause of CMT type 1A (CMT1A) and HNPP. The importance of a sensitive and rapid method for identifying the CMT1A duplication and HNPP deletion is being emphasized. In the present study, we established a molecular diagnostic method for the CMT1A duplication and HNPP deletion based on hexaplex PCR of 6 microsatellite markers (D17S921, D17S9B, D17S9A, D17S918, D17S4A and D17S2230). The method is highly time-, cost- and sample-saving because the six markers are amplified by a single PCR reaction and resolved with a single capillary in 3 h. Several statistical and forensic estimates indicated that most of these markers are likely to be useful for diagnosing the peripheral neuropathies. Reproducibility, as determined by concordance between independent tests, was estimated to be 100%. The likelihood that genotypes of all six markers are homozygous in randomly selected individuals was calculated to be 1.6 x 10(-4) which indicates that the statistical error rate for this diagnosis of HNPP deletion is only 0.016%.
Choi
Byung-Ok
BO
Department of Biological Science, Kongju National University, Gongju 314-701, Korea.
Kim
Joonki
J
Lee
Kyung Lyong
KL
Yu
Jin Seok
JS
Hwang
Jung Hee
JH
Chung
Ki Wha
KW
D000483:Alleles; D002607:Charcot-Marie-Tooth Disease / Q000175:diagnosis* / Q000235:genetics*; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage; D020440:Gene Duplication; D005787:Gene Frequency; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D010375:Pedigree; D010523:Peripheral Nervous System Diseases / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics