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Universal fluorescent multiplex PCR and capillary electrophoresis for evaluation of gene conversion between SMN1 and SMN2 in spinal muscular atrophy

Analytical and Bioanalytical Chemistry volume 397, pages 2375–2383 (2010)Cite this article

Abstract

We have developed a capillary electrophoresis (CE) method with universal fluorescent multiplex PCR to simultaneously detect the SMN1 and SMN2 genes in exons 7 and 8. Spinal muscular atrophy (SMA) is a very frequent inherited disease caused by the absence of the SMN1 gene in approximately 94% of patients. Those patients have deletion of the SMN1 gene or gene conversion between SMN1 and SMN2. However, most methods only focus on the analysis of whole gene deletion, and ignore gene conversion. Simultaneous quantification of SMN1 and SMN2 in exons 7 and 8 is a good strategy for estimating SMN1 deletion or SMN1 to SMN2 gene conversion. This study established a CE separation allowing differentiation of all copy ratios of SMN1 to SMN2 in exons 7 and 8. Among 212 detected individuals, there were 23 SMA patients, 45 carriers, and 144 normal subjects. Three individuals had different ratios of SMN1 to SMN2 in two exons, including an SMA patient having two SMN2 copies in exon 7 but one SMN1 copy in exon 8. This method could provide more information about SMN1 deletion or SMN1 to SMN2 gene conversion for SMA genotyping and diagnosis.

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Acknowledgements

We gratefully acknowledge the support of the National Science Council of Taiwan and the Center of Excellence for Environmental Medicine (KMU-EM-98-1-2) of Kaohsiung Medical University in funding this work. We also express our cordial gratitude to Sun’s KMU-SMA fund for partial financial support of this work.

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Affiliations

  1. School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Chun-Chi Wang & Shou-Mei Wu

  2. Graduate Institute of Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Yuh-Jyh Jong & Jan-Gowth Chang

  3. Department of Laboratory Medicine, Kaohsiung Medical University Hospital, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Yuh-Jyh Jong & Jan-Gowth Chang

  4. Department of Pediatrics, Kaohsiung Medical University Hospital, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Yuh-Jyh Jong

  5. Center of Excellence for Environmental Medicine, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Jan-Gowth Chang & Shou-Mei Wu

  6. Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan

    Yen-Ling Chen

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  1. Chun-Chi Wang
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  2. Yuh-Jyh Jong
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  3. Jan-Gowth Chang
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  4. Yen-Ling Chen
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  5. Shou-Mei Wu
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Correspondence to Shou-Mei Wu.

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Wang, CC., Jong, YJ., Chang, JG. et al. Universal fluorescent multiplex PCR and capillary electrophoresis for evaluation of gene conversion between SMN1 and SMN2 in spinal muscular atrophy. Anal Bioanal Chem 397, 2375–2383 (2010). https://doi.org/10.1007/s00216-010-3761-1

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  • DOI: https://doi.org/10.1007/s00216-010-3761-1

Keywords

  • Gene conversion
  • Spinal muscular atrophy
  • SMN1/SMN2
  • Universal fluorescent multiplex PCR
  • Capillary electrophoresis