Carrier Detection in Duchenne Muscular Dystrophy Using Molecular Methods

Overview

Journal Indian J Med Res

Specialty General Medicine

Date 2013 Jul 16

PMID 23852291

Citations 12

Authors

S M Sakthivel Murugan

C Arthi

N Thilothammal

B R Lakshmi

Affiliations

Soon will be listed here.

Abstract

Background & Objectives: Duchenne and Becker muscular dystrophies are X-linked allelic disorders which are caused by mutations in the DMD gene. Carrier analysis in DMD is complicated due to the heterozygous nature of the X chromosome. Several techniques have been tried for carrier analysis in families where the mutation is identified including quantitative multiplex PCR (qmPCR), Southern blot, and now multiplex ligation-dependent probe amplification (MLPA). Linkage analysis is used in cases without identifiable mutations. The present study was undertaken to determine the status of probable carriers in families where the DMD deletion/duplication has been identified for the affected index cases.

Methods: Carrier status was present in 150 probable carriers from 110 apparently unrelated families where the patients' mutations were known. Of these 110 families, 100 were deletions, 9 duplications and 1 point mutation. Multiplex ligation-dependent probe amplification (MLPA) was used to assess the copy number changes and direct sequencing was used for the case with the point mutation.

Results: Of the 150 cases, 49 were found to be carriers. Among the sporadic cases, it was observed that the rate of de novo mutations was very high (71%) as compared to the hereditary cases (29%), which was higher than the calculated rate (30%). It was observed that this difference was more apparent in deletion mutations than in duplications.

Interpretation & Conclusions: Identifying the DMD carrier rates in the families with unidentified deletions and duplications and where the causative mutation could be small insertions/deletions or point mutations could throw more light into this observation. MLPA was found to be useful in detecting copy number changes in DMD carriers and this could be the method of choice for DMD carrier analysis, when the mutation is detected in the affected child.

Citing Articles

Pragmatic Neurorehabilitation Approach for Improving Quality of Life in Duchenne Muscular Dystrophy: A Case Report.

Nangliya R, Sasun A, Samal S Cureus. 2024; 16(3):e56315.

PMID: 38629006 PMC: 11020631. DOI: 10.7759/cureus.56315.

Preliminary Results of a Genetic Study of Children with Duchenne Myodystrophy in Aktobe, Kazakhstan.

Umurzakova A, Ayaganov D, Mannapova A, Dzhumasheva B, Dzhaksybayeva A Arch Razi Inst. 2023; 78(3):949-954.

PMID: 38028863 PMC: 10657926. DOI: 10.22092/ARI.2022.360050.2534.

Mutation spectrum analysis of gene in Indonesian Duchenne and Becker muscular dystrophy patients.

Dwianingsih E, Iskandar K, Hapsara S, Ping Liu C, Malueka R, Gunadi F1000Res. 2023; 11:148.

PMID: 38009102 PMC: 10668572. DOI: 10.12688/f1000research.73476.3.

Results of a genetic study of children with Duchenne myodystrophy in Kazakhstan.

Umurzakova A, Ayaganov D, Nurgaliyeva R J Neurosci Rural Pract. 2023; 14(2):389-390.

PMID: 37181168 PMC: 10174136. DOI: 10.25259/JNRP_55_2022.

Diagnosing Muscular Dystrophies: Comparison of Techniques and Their Cost Effectiveness: A Multi-institutional Study.

Srivastava P, Malhotra K, Husain N, Malhotra H, Kulshreshtha D, Anand A J Neurosci Rural Pract. 2020; 11(3):420-429.

PMID: 32753807 PMC: 7394627. DOI: 10.1055/s-0040-1713301.

References

Chaturvedi L, Mittal R, Srivastava S, Mukherjee M, Mittal B . Analysis of dinucleotide repeat loci of dystrophin gene for carrier detection, germline mosaicism and de novo mutations in Duchenne muscular dystrophy. Clin Genet. 2000; 58(3):234-6. DOI: 10.1034/j.1399-0004.2000.580312.x. View

EMERY A . Population frequencies of inherited neuromuscular diseases--a world survey. Neuromuscul Disord. 1991; 1(1):19-29. DOI: 10.1016/0960-8966(91)90039-u. View

Miller S, Dykes D, Polesky H . A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988; 16(3):1215. PMC: 334765. DOI: 10.1093/nar/16.3.1215. View

Murugan S, Chandramohan A, Lakshmi B . Use of multiplex ligation-dependent probe amplification (MLPA) for Duchenne muscular dystrophy (DMD) gene mutation analysis. Indian J Med Res. 2010; 132:303-11. View

Gatta V, Scarciolla O, Gaspari A, Palka C, De Angelis M, Di Muzio A . Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA). Hum Genet. 2005; 117(1):92-8. DOI: 10.1007/s00439-005-1270-7. View

Peier A, Caskey C, Richards C, Shaffer L . DMD-specific FISH probes are diagnostically useful in the detection of female carriers of DMD gene deletions. Neurology. 1997; 48(6):1633-8. DOI: 10.1212/wnl.48.6.1633. View

Mukherjee M, Chaturvedi L, Srivastava S, Mittal R, Mittal B . De novo mutations in sporadic deletional Duchenne muscular dystrophy (DMD) cases. Exp Mol Med. 2003; 35(2):113-7. DOI: 10.1038/emm.2003.16. View

Ligon A, Kashork C, Richards C, Shaffer L . Identification of female carriers for Duchenne and Becker muscular dystrophies using a FISH-based approach. Eur J Hum Genet. 2000; 8(4):293-8. DOI: 10.1038/sj.ejhg.5200450. View

Alcantara M, Villarreal M, Del Castillo V, Gutierrez G, Saldana Y, Maulen I . High frequency of de novo deletions in Mexican Duchenne and Becker muscular dystrophy patients. Implications for genetic counseling. Clin Genet. 1999; 55(5):376-80. DOI: 10.1034/j.1399-0004.1999.550514.x. View

10.

Yau S, Bobrow M, Mathew C, Abbs S . Accurate diagnosis of carriers of deletions and duplications in Duchenne/Becker muscular dystrophy by fluorescent dosage analysis. J Med Genet. 1996; 33(7):550-8. PMC: 1050661. DOI: 10.1136/jmg.33.7.550. View

11.

Janssen B, Hartmann C, Scholz V, Jauch A, Zschocke J . MLPA analysis for the detection of deletions, duplications and complex rearrangements in the dystrophin gene: potential and pitfalls. Neurogenetics. 2005; 6(1):29-35. DOI: 10.1007/s10048-004-0204-1. View

12.

Bakker E, Bonten E, De Lange L, Veenema H, Majoor-Krakauer D, Hofker M . DNA probe analysis for carrier detection and prenatal diagnosis of Duchenne muscular dystrophy: a standard diagnostic procedure. J Med Genet. 1986; 23(6):573-80. PMC: 1049839. DOI: 10.1136/jmg.23.6.573. View

13.

Clemens P, Fenwick R, Chamberlain J, Gibbs R, de Andrade M, Chakraborty R . Carrier detection and prenatal diagnosis in Duchenne and Becker muscular dystrophy families, using dinucleotide repeat polymorphisms. Am J Hum Genet. 1991; 49(5):951-60. PMC: 1683265. View

14.

Dubowitz V . The female carrier of Duchenne muscular dystrophy. Br Med J (Clin Res Ed). 1982; 284(6327):1423-4. PMC: 1498325. DOI: 10.1136/bmj.284.6327.1423. View

15.

Abbs S, Tuffery-Giraud S, Bakker E, Ferlini A, Sejersen T, Mueller C . Best practice guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies. Neuromuscul Disord. 2010; 20(6):422-7. DOI: 10.1016/j.nmd.2010.04.005. View

16.

Carsana A, Frisso G, Tremolaterra M, Ricci E, De Rasmo D, Salvatore F . A larger spectrum of intragenic short tandem repeats improves linkage analysis and localization of intragenic recombination detection in the dystrophin gene: an analysis of 93 families from southern Italy. J Mol Diagn. 2007; 9(1):64-9. PMC: 1867430. DOI: 10.2353/jmoldx.2007.060056. View

17.

Kumari D, Mital A, Gupta M, Goyle S . Deletion analysis of the dystrophin gene in Duchenne and Becker muscular dystrophy patients: use in carrier diagnosis. Neurol India. 2003; 51(2):223-6. View

18.

Taylor P, Maroulis S, Mullan G, Pedersen R, Baumli A, Elakis G . Measurement of the clinical utility of a combined mutation detection protocol in carriers of Duchenne and Becker muscular dystrophy. J Med Genet. 2007; 44(6):368-72. PMC: 2740880. DOI: 10.1136/jmg.2006.047464. View

19.

Panigrahi I, Mittal B . Carrier detection and prenatal diagnosis in Duchenne/Becker muscular dystrophy. Indian Pediatr. 2001; 38(6):631-9. View

20.

Sinha S, Mishra S, Singh V, Mittal R, Mittal B . High frequency of new mutations in North Indian Duchenne/Becker muscular dystrophy patients. Clin Genet. 1996; 50(5):327-31. DOI: 10.1111/j.1399-0004.1996.tb02383.x. View