title: Diagnosis of Duchenne and Becker muscular dystrophies by polymerase chain reaction. A multicenter study.
abstract: OBJECTIVE--To assess the efficiency, reliability, and ease of use of DNA diagnosis for Duchenne and Becker muscular dystrophies (DMD/BMD) using the polymerase chain reaction (PCR). DESIGN--DNA from the patients was screened for deletion mutations using multiplex PCR, and the results were compared with those obtained by Southern blot analysis. The PCR multiplex reaction detects nine specific "hot-spot" exons in the dystrophin gene while the Southern analysis detects 66 specific dystrophin gene restriction fragments. The multiplex reaction requires 50-fold less DNA than Southern analysis and thus is considerably more sensitive. SETTING--Fourteen university-affiliated and private genetic disease diagnostic laboratories. PATIENTS--Male patients with clinical signs of DMD/BMD. Cases were selected for analysis randomly, without knowledge of whether a deletion was present within the dystrophin gene. MAIN OUTCOME MEASURES--The percentage of cases that were detectable by multiplex PCR in comparison with Southern analysis, the frequency, extent, and location of the detected deletion mutations. In some cases, duplication mutations were monitored. RESULTS--The accuracy of a single PCR multiplex amplification (nine exons) was compared with Southern analysis with 10 cDNA probes that cover the full length of the gene. The multiplex PCR analytic method detected 82% of those deletions detected by Southern analysis methods. In one of 745 analyses, the multiplex method suggested a single exon deletion, which was not confirmed by Southern analysis, representing a false-positive rate of 0.013%. CONCLUSIONS--Multiplex PCR represents a sensitive and accurate method for deletion detection of 46% of all cases of DMD/BMD. The method requires 1 day for analysis, is easy to perform, and does not use radioactive tracers. As such, multiplex PCR represents an efficient and rapid method for prenatal or postnatal diagnosis of DMD/BMD.
authors.forename: J S
authors.initials: JS
authors.lastname: Chamberlain
authors.forename: J R
authors.initials: JR
authors.lastname: Chamberlain
authors.forename: R G
authors.initials: RG
authors.lastname: Fenwick
authors.forename: P A
authors.initials: PA
authors.lastname: Ward
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
authors.forename: L S
authors.initials: LS
authors.lastname: Dimnik
authors.forename: N T
authors.initials: NT
authors.lastname: Bech-Hansen
authors.forename: D I
authors.initials: DI
authors.lastname: Hoar
authors.forename: S
authors.initials: S
authors.lastname: Richards
authors.forename: A E
authors.initials: AE
authors.lastname: Covone
authors.forename: R
authors.initials: R
authors.lastname: Govanni
authors.forename: S
authors.initials: S
authors.lastname: Abbs
authors.forename: D R
authors.initials: DR
authors.lastname: Bentley
authors.forename: M
authors.initials: M
authors.lastname: Bobrow
authors.forename: G
authors.initials: G
authors.lastname: Rysiecki
authors.forename: P N
authors.initials: PN
authors.lastname: Ray
authors.forename: C
authors.initials: C
authors.lastname: Boileau
authors.forename: C
authors.initials: C
authors.lastname: Junien
authors.forename: C
authors.initials: C
authors.lastname: Boehm
authors.forename: V L
authors.initials: VL
authors.lastname: Venne
authors.forename: F K
authors.initials: FK
authors.lastname: Fujmura
authors.forename: I
authors.initials: I
authors.lastname: Spiga
authors.forename: M
authors.initials: M
authors.lastname: Ferrari
authors.forename: S
authors.initials: S
authors.lastname: Tedeschi
authors.forename: E
authors.initials: E
authors.lastname: Bakker
authors.forename: A L
authors.initials: AL
authors.lastname: Kneppers
authors.forename: G J
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: K
authors.initials: K
authors.lastname: Jain
authors.forename: E
authors.initials: E
authors.lastname: Spector
authors.forename: B
authors.initials: B
authors.lastname: Crandall
authors.forename: A
authors.initials: A
authors.lastname: Kiuru
authors.forename: M L
authors.initials: ML
authors.lastname: Savontaus
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
authors.forename: J S
authors.initials: JS
authors.lastname: Chamberlain
authors.forename: J R
authors.initials: JR
authors.lastname: Chamberlain
authors.forename: G
authors.initials: G
authors.lastname: Rysiecki
mesh_terms: D015139:Blotting, Southern; D002872:Chromosome Deletion; D004247:DNA / Q000032:analysis; D006801:Humans; D008297:Male; D009136:Muscular Dystrophies / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011446:Prospective Studies
chemical_list: D004247:DNA