title: Evaluation of an in-house protocol for prenatal molecular diagnosis of SMA in Chinese.
abstract: BACKGROUND
  Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration of the anterior horn of the spinal cord, leading to symmetric muscle weakness and atrophy. About 95% of SMA patients have homozygous loss of SMN1 which can be detected by conventional PCR-RFLP testing. However, the method cannot distinguish heterozygous healthy carriers. A quantitative method named multiple ligation-dependent probe amplification (MLPA) was introduced in our protocol for prenatal molecular diagnosis of SMA in our laboratory.
  
  
  METHODS
  DNA was extracted from amniotic fluid cells of 13 fetuses from 11 Chinese SMA families. STR profiling was carried out to evaluate the contamination of amniotic DNA by maternal genomic DNA. Three methods, PCR-RFLP, allele-specific PCR and MLPA, were used to analyze SMN1 exon 7 of amniotic DNA.
  
  
  RESULTS
  There was no contamination of amniotic DNA by maternal genomic DNA. In conventional PCR-RFLP, allele-specific PCR, and MLPA, homozygous loss of SMN1 was observed in 4 fetuses. Among the remaining 9 fetuses, 6 with 1 copy of SMN1 and 3 with 2 copies of SMN1 were showed by MLPA.
  
  
  CONCLUSIONS
  This in-house protocol was reliable and efficient for prenatal molecular diagnosis of SMA.
authors.affiliation: Research Center for Molecular Diagnosis of Genetic Diseases, Fuzhou General Hospital, Fuzhou City, Fujian Province, 350025 PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zeng
authors.forename: Fenghua
authors.initials: F
authors.lastname: Lan
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Deng
authors.forename: Longfeng
authors.initials: L
authors.lastname: Ke
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Tu
authors.forename: Lianghu
authors.initials: L
authors.lastname: Huang
authors.forename: Dezhu
authors.initials: D
authors.lastname: Zheng
authors.forename: Zhongyong
authors.initials: Z
authors.lastname: Zhu
mesh_terms: D000328:Adult; D000483:Alleles; D000653:Amniotic Fluid / Q000737:chemistry; D002681:China; D004247:DNA / Q000032:analysis / Q000235:genetics; D005091:Exons; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D012150:Polymorphism, Restriction Fragment Length; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014897:Spinal Muscular Atrophies of Childhood / Q000175:diagnosis*; D055815:Young Adult
chemical_list: D004247:DNA