Skip to main content
SpringerLink
Log in

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

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Feldkötter M, Schwarzer V, Wirth R, Wienker TF, Wirth B (2002) Am J Hum Genet 70:358–368

    Article  Google Scholar 

  2. Melki J, Lefebvre S, Burglen L, Burlet P, Clermont O, Millasseau P, Reboullet S, Bénichou B, Zeviani M, Paslier DL, Cohen D, Weissenbach J, Munnich A (1994) Science 264:1474–1477

    Article  CAS  Google Scholar 

  3. Ogino S, Wilson RB (2002) Hum Genet 111:477–500

    Article  CAS  Google Scholar 

  4. Zerres K, Rudnik-Schöneborn S (1995) Arch Neurol 52:518–523

    CAS  Google Scholar 

  5. Monani UR, Lorson CL, Parsons DW, Prior TW, Androphy EJ, Burghes AHM, McPherson JD (1999) Hum Mol Genet 8:1177–1183

    Article  CAS  Google Scholar 

  6. Lunn MR, Wang CH (2008) Lancet 371:2120–2133

    Article  Google Scholar 

  7. Wirth B, Herz M, Wetter A, Moskau S, Hahnen E, Rudnik-Schöneborn S, Wienker T, Zerres K (1999) Am J Hum Genet 64:1340–1356

    Article  CAS  Google Scholar 

  8. Lefebvre S, Burglen L, Reboullet S, Clermont O, Burlet P, Viollet L, Benichou B, Cruaud C, Millasseau P, Zeviani M, Paslier DL, Frézal J, Cohen D, Weissenbach J, Munnich A, Melki J (1995) Cell 80:155–165

    Article  CAS  Google Scholar 

  9. McAndrew PE, Parsons DW, Simard LR, Rochette C, Ray PN, Mendell JR, Prior TW, Burphes AHM (1997) Am J Hum Genet 60:1411–1422

    Article  CAS  Google Scholar 

  10. Burphes AHM (1997) Am J Hum Genet 61:9–15

    Article  Google Scholar 

  11. Hahnen E, Sćhonling J, Rudnik-Schöneborn S, Zerres K, Wirth B (1996) Am J Hum Genet 59:1057–1065

    CAS  Google Scholar 

  12. Campbell L, Potter A, Ignatius J, Davies K (1997) Am J Hum Genet 61:40–50

    Article  CAS  Google Scholar 

  13. DiDonato CJ, Ingraham SE, Mendell JR, Prior TW, Lenard S, Moxley RT, Florence J, Burghes AHM (1997) Ann Neurol 41:230–237

    Article  CAS  Google Scholar 

  14. Ogino S, Gao S, Leonard DG, Paessler M, Wilson RB (2003) Eur J Hum Genet 11:275–277

    Article  CAS  Google Scholar 

  15. Mazzei R, Gambardella A, Conforti FL, Magariello A, Patitucci A, Gabriele AL, Sprovieri T, Labate A, Valentino P, Bono F, Bonavita S, Zappia M, Muglia M, Quattrone A (2004) Acta Neurol Scand 109:151–154

    Article  CAS  Google Scholar 

  16. Gérard B, Ginet N, Matthijs G, Evrard P, Baumann C, Da Silva F, Gérard-Blanluet M, Mayer M, Grandchamp B, Elion J (2000) Hum Mutat 16:253–263

    Article  Google Scholar 

  17. Anhuf D, Eggermann T, Rudnik-Schöneborn S, Zerres K (2003) Hum Mutat 22:74–78

    Article  CAS  Google Scholar 

  18. Sutomo R, Akutsu T, Takeshima Y, Nishio H, Sadewa AH, Harada Y, Matsuo M (2002) Am J Med Genet 113:225–226

    Article  Google Scholar 

  19. Su YN, Hung CC, Li H, Lee CN, Cheng WF, Tsao PN, Chang MC, Yu CL, Hsieh WS, Lin WL, Hsu SM (2005) Hum Mutat 25:460–467

    Article  CAS  Google Scholar 

  20. Hung CC, Lee CN, Chen CP, Jong YJ, Chen CA, Cheng WF, Lin WL, Su YL (2005) Anal Chem 77:6960–6968

    Article  CAS  Google Scholar 

  21. Hung CC, Chien SC, Lin CY, Chang CH, Chang YF, Jong YJ, Hsieh ST, Hsieh WS, Liu MS, Lin WL, Lee CN, Su YN (2007) Electrophoresis 28:2826–2834

    Article  CAS  Google Scholar 

  22. Kao HY, Su YN, Liao HK, Liu MS, Chen YJ (2006) Clin Chem 52:361–369

    Article  CAS  Google Scholar 

  23. Huang CH, Chang YY, Chen CH, Kuo YS, Hwu WL, Gerdes T, Ko TM (2007) Genet Med 9:241–248

    Article  Google Scholar 

  24. Zeng J, Lan F, Deng X, Ke L, Tu X, Huang L, Zheng D, Zhu Z (2008) Clin Chim Acta 398:78–81

    Article  CAS  Google Scholar 

  25. Wang CC, Chang JG, Ferrance J, Chen HY, You CY, Chang YF, Jong YJ, Wu SM, Yeh CH (2008) Electrophoresis 29:2904–2911

    Article  CAS  Google Scholar 

  26. Wang CC, Chang JG, Jong YJ, Wu SM (2009) Electrophoresis 30:1102–1110

    Article  CAS  Google Scholar 

  27. Song L, Liu T, Liang D, Fang D, Chu B (2001) Electrophoresis 22:2688–2698

    Google Scholar 

  28. Wang Y, Liang D, Hao J, Fang D, Chu B (2002) Electrophoresis 23:1460–1466

    Article  CAS  Google Scholar 

  29. Kan CW, Barron AE (2003) Electrophoresis 24:55–62

    Article  CAS  Google Scholar 

  30. Wang Y, Liang D, Ying Q, Chu B (2005) Electrophoresis 26:126–136

    Article  Google Scholar 

  31. Chou LS, Gedge F, Lyon E (2005) J Mol Diagn 7:111–120

    CAS  Google Scholar 

  32. Li-Sucholeiki XC, Hu G, Perls T, Tomita-Mitchell A, Thilly WG (2005) Electrophoresis 26:2531–2538

    Article  CAS  Google Scholar 

  33. DiMatteo D, Callahan S, Kmiec EB (2008) Exp Cell Res 314:878–886

    Article  CAS  Google Scholar 

Download references

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.

Author information

Authors and 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

Authors
  1. Chun-Chi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuh-Jyh Jong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan-Gowth Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yen-Ling Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shou-Mei Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shou-Mei Wu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

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

Keywords

Search

Navigation