Allele-specific all-or-none PCR product diagnostic strategy for CMT 1A and HNPP

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J Chin Med Assoc. 2006 Feb

Allele-specific all-or-none PCR product diagnostic strategy for

Charcot-Marie-Tooth 1A and hereditary neuropathy with liability to

pressure palsies.

Lin KP, Chou CH, Lee HY, Soong BW.

The Neurological Institute, Taipei Veterans General Hospital, Taiwan,

ROC.

BACKGROUND: We designed allele-specific primers to amplify genomic

DNA of patients with Charcot-Marie-Tooth 1A (CMT1A) and hereditary

neuropathy with liability to pressure palsies (HNPP).

METHODS: Genomic DNA analysis was performed on 40 unrelated CMT1A

duplication patients, 25 unrelated HNPP deletion patients, and 50

unaffected control individuals. The CMT1A and HNPP patients had

previously been identified with microsatellite mapping.

RESULTS: Amplification products came to 3.6 kb in length from the

normal proximal CMT1A repeated segment on chromosome 17p11.2

(proximal CMT1A-REP), 3.57 kb from the normal distal CMT1A repeated

segment on chromosome 17p11.2 (distal CMT1A-REP), 3.6 kb from HNPP

patients, and 3.58 kb from CMT1A patients. We could identify the

mutations by means of agarose gel electrophoresis after polymerase

chain reaction (PCR) amplification without restriction enzyme

digestion from 33 of the 40 CMT1A and 19 of the 25 HNPP samples.

CONCLUSION: Stringently specific primers were used to overcome the

problem of nonspecific amplification and provide a rapid, all-or-none

PCR product and efficient screening test for CMT1A and HNPP.