PMID- 15988805
OWN - NLM
STAT- MEDLINE
DCOM- 20050817
LR  - 20181113
IS  - 0513-5796 (Print)
IS  - 1976-2437 (Electronic)
IS  - 0513-5796 (Linking)
VI  - 46
IP  - 3
DP  - 2005 Jun 30
TI  - Effectiveness of real-time quantitative PCR compare to repeat PCR for the 
      diagnosis of Charcot-Marie-Tooth Type 1A and hereditary neuropathy with liability 
      to pressure palsies.
PG  - 347-52
AB  - The majority of cases of Charcot-Marie-Tooth type 1A (CMT1A) and of hereditary 
      neuropathy with a liability to pressure palsies (HNPP) are the result of 
      heterozygosity for the duplication or deletion of peripheral myelin protein 22 
      gene (PMP22) on 17p11.2. Southern blots, pulsed-field gel electrophoresis (PFGE), 
      fluorescence in situ hybridization (FISH) and polymorphic marker analysis are 
      currently used diagnostic methods. But they are time-consuming, labor-intensive 
      and have some significant limitations. We describe a rapid real- time 
      quantitative PCR method for determining gene copy number for the identification 
      of DNA duplication or deletion occurring in CMT1A or HNPP and compare the results 
      obtained with REP-PCR. Six patients with CMT1A and 14 patients with HNPP 
      [confirmed by Repeat (REP)-PCR], and 16 patients with suspicious CMT1A and 13 
      patients with suspicious HNPP [negative REP-PCR], and 15 normal controls were 
      studied. We performed REP-PCR, which amplified a 3.6 Kb region (including a 1.7Kb 
      recombination hotspot), using specific CMT1A-REP and real-time quantitative PCR 
      on the LightCycler system. Using a comparative threshold cycle (Ct) method and 
      beta -globin as a reference gene, the gene copy number of the PMP22 gene was 
      quantified. The PMP22 duplication ratio ranged from 1.35 to 1.74, and the PMP22 
      deletion ratio from 0.41 to 0.53. The PMP22 ratio in normal controls ranged from 
      0.81 to 1.12. All 6 patients with CMT1A and 14 patients with HNPP confirmed by 
      REP-PCR were positive by real-time quantitative PCR. Among the 16 suspicious 
      CMT1A and 13 suspicious HNPP with negative REP-PCR, 2 and 4 samples, 
      respectively, were positive by real-time quantitative PCR. Real-time quantitative 
      PCR is a more sensitive and more accurate method than REP-PCR for the detection 
      of PMP22 duplications or deletions, and it is also faster and easier than 
      currently available methods. Therefore, we believe that the real-time 
      quantitative method is useful for diagnosing CMT1A and HNPP.
FAU - Choi, Jong Rak
AU  - Choi JR
AD  - Departments of Laboratory Medicine, Yonsei University College of Medicine, 134 
      Shinchon-dong, Seodaemoon-gu, Seoul 120-752, Korea.
FAU - Lee, Woon Hyoung
AU  - Lee WH
FAU - Sunwoo, Il Nam
AU  - Sunwoo IN
FAU - Lee, Eun Kyung
AU  - Lee EK
FAU - Lee, Chang Hoon
AU  - Lee CH
FAU - Lim, Jong Baeck
AU  - Lim JB
LA  - eng
PT  - Clinical Trial
PT  - Comparative Study
PT  - Controlled Clinical Trial
PT  - Journal Article
PL  - Korea (South)
TA  - Yonsei Med J
JT  - Yonsei medical journal
JID - 0414003
SB  - IM
MH  - Charcot-Marie-Tooth Disease/*diagnosis/*genetics
MH  - Gene Dosage
MH  - Genetic Testing/methods
MH  - Hereditary Sensory and Motor Neuropathy/*diagnosis/*genetics
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
PMC - PMC2815810
EDAT- 2005/07/01 09:00
MHDA- 2005/08/18 09:00
PMCR- 2005/06/30
CRDT- 2005/07/01 09:00
PHST- 2005/07/01 09:00 [pubmed]
PHST- 2005/08/18 09:00 [medline]
PHST- 2005/07/01 09:00 [entrez]
PHST- 2005/06/30 00:00 [pmc-release]
AID - 200506347 [pii]
AID - 10.3349/ymj.2005.46.3.347 [doi]
PST - ppublish
SO  - Yonsei Med J. 2005 Jun 30;46(3):347-52. doi: 10.3349/ymj.2005.46.3.347.