title: New multiplex real-time PCR approach to detect gene mutations for spinal muscular atrophy.
abstract: BACKGROUND
  Spinal muscular atrophy (SMA) is the most common autosomal recessive disease in children, and the diagnosis is complicated and difficult, especially at early stage. Early diagnosis of SMA is able to improve the outcome of SMA patients. In our study, Real-time PCR was developed to measure the gene mutation or deletion of key genes for SMA and to further analyse genotype-phenotype correlation.
  
  
  METHODS
  The multiple real-time PCR for detecting the mutations of survival of motor neuron (SMN), apoptosis inhibitory protein (NAIP) and general transcription factor IIH, polypeptide 2 gene (GTF2H2) was established and confirmed by DNA sequencing and multiplex ligation-dependent probe amplification (MLPA). The diagnosis and prognosis of 141 hospitalized children, 100 normal children and further 2000 cases of dry blood spot (DBS) samples were analysed by this multiple real-time PCR.
  
  
  RESULTS
  The multiple real-time PCR was established and the accuracy of it to detect the mutations of SMN, NAIP and GTF2H2 was at least 98.8 % comparing with DNA sequencing and MLPA. Among 141 limb movement disorders children, 75 cases were SMA. 71 cases of SMA (94.67 %) were with SMN c.840 mutation, 9 cases (12 %) with NAIP deletion and 3 cases (4 %) with GTF2H2 deletion. The multiple real-time PCR was able to diagnose and predict the prognosis of SMA patients. Simultaneously, the real-time PCR was applied to detect trace DNA from DBS and able to make an early diagnosis of SMA.
  
  
  CONCLUSIONS
  The clinical and molecular characteristics of SMA in Southwest of China were presented. Our work provides a novel way for detecting SMA in children by using real-time PCR and the potential usage in newborn screening for early diagnosis of SMA.
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Zhidai
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Center for Clinical Laboratory, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Penghui
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: He
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Liu
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Shi
authors.initials: S
authors.lastname: Tang
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Xinbin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Tan
authors.affiliation: Department of Health Statistics, School of Public Health, Chongqing Medical University, Yuzhong District, Chongqing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Peng
authors.affiliation: Department of Neurology, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Department of Neurology, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China.
authors.forename: Siqi
authors.initials: S
authors.lastname: Hong
authors.affiliation: Center for Clinical Molecular Medicine, Children's Hospital, Chongqing Medical University, 136 Zhongshan Er Road, Yuzhong District, Chongqing, 400014, China. 2660622115@qq.com.|Ministry of Education Key Laboratory of Development and Disorders, Children's Hospital, Chongqing Medical University, Yuzhong District, Chongqing, China. 2660622115@qq.com.|Key Laboratory of Pediatrics in Chongqing, Children's Hospital, Chongqing Medical University, Chongqing, China. 2660622115@qq.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Zou
mesh_terms: D002648:Child; D002675:Child, Preschool; D002681:China; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D009134:Muscular Atrophy, Spinal / Q000175:diagnosis* / Q000235:genetics*; D015997:Neonatal Screening / Q000379:methods; D009419:Nerve Tissue Proteins / Q000235:genetics; D051616:Neuronal Apoptosis-Inhibitory Protein / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D055533:Survival of Motor Neuron 1 Protein / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
chemical_list: C496800:NAIP protein, human; D009419:Nerve Tissue Proteins; D051616:Neuronal Apoptosis-Inhibitory Protein; D055533:Survival of Motor Neuron 1 Protein; D014157:Transcription Factors
keywords: Multiplex real-time PCR; Newborn screening; Spinal muscular atrophy