title: Development of a comprehensive real-time PCR assay for dystrophin gene analysis and prenatal diagnosis of Chinese families.
abstract: BACKGROUND
  To develop a comprehensive method to analyze deletions or duplications of the dystrophin gene in both patients and carriers of Duchenne muscular dystrophy (DMD), likewise applied to prenatal diagnosis.
  
  
  METHODS
  A total of thirty Chinese families were recruited, composed of 29 DMD affected males and 38 female relatives containing four pregnant women. Deletions were previously screened by multiplex PCR. A comprehensive real-time PCR assay using SYBR Green I dye was performed for the initial detection of duplications in patients with a seven-exon primer set, carrier detection for female relatives and prenatal diagnosis for the 4 of them. The results were later confirmed by multiple ligation-dependent probe amplification (MLPA) and linkage analysis.
  
  
  RESULTS
  Three out of 4 duplications were first discovered by real-time PCR. Carrier status was ascertained in 22 and rejected in the remaining sixteen female relatives. Furthermore, 4 fetuses were diagnosed as two normal females, one normal male and one female carrier, respectively.
  
  
  CONCLUSIONS
  Our real-time PCR assay is useful in duplication screen with a detection rate of >70%, as well as rapid and reliable in both carrier detection and prenatal diagnosis of DMD families with known deletions and duplications.
authors.affiliation: Department of Genetics, School of Medicine, Zhejiang University, 866 Yuhangtang Road, Hangzhou, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.forename: Shaoji
authors.initials: S
authors.lastname: Liu
authors.forename: Tianying
authors.initials: T
authors.lastname: Wei
authors.forename: Jing
authors.initials: J
authors.lastname: Yong
authors.forename: Yuchan
authors.initials: Y
authors.lastname: Mao
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Lu
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Xie
authors.forename: Qing
authors.initials: Q
authors.lastname: Ke
authors.forename: Fan
authors.initials: F
authors.lastname: Jin
authors.forename: Ming
authors.initials: M
authors.lastname: Qi
mesh_terms: D000328:Adult; D044466:Asian People; D016189:Dystrophin / Q000235:genetics*; D005091:Exons; D005260:Female; D005333:Fetus; D017353:Gene Deletion; D018628:Gene Dosage; D020440:Gene Duplication; D008040:Genetic Linkage; D006579:Heterozygote; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis* / Q000208:ethnology / Q000235:genetics; D010375:Pedigree; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D016189:Dystrophin
keywords: Carrier detection; Deletion; Duchenne muscular dystrophy; Duplication; Prenatal diagnosis; Real-time PCR