E-mail ID : info@iamg.in
 
 
 
 

Online Submission
Click Here For Online Submission
Instructions for authors

Genetic Clinics
Editorial board

Get Our Newsletter
Subscribe

Send Your Feedback
Feedback Form

About Us
IAMG

Abstract

October - December 2023 | Vol. 16 | Issue 4 | 14-19
Validation of MLPA-detected Single Exon Deletion of the DMD Gene by Multiplex PCR
Haseena Sait, Shubha R Phadke
Department of Medical Genetics, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
Address for Correspondence Email: shubharaophadke@gmail.com
 
Abstract
Multiplex ligation-dependent probe amplification (MLPA) is the most widely used technique to detect deletions and duplications of the DMD (dystrophin) gene that constitute two third of the cases of Duchenne (DMD)/ Becker muscular dystrophy (BMD). However, MLPA can yield false positive results for single exon deletions owing to the presence of single nucleotide polymorphisms (SNPs) at the ligation site. This study aimed to validate single exon deletions detected by MLPA in DMD/BMD individuals by multiplex polymerase chain reaction (PCR). The data of patients with DMD/BMD diagnosed by MLPA between January 2016 to February 2021 was collated and those with apparent single exon deletions were recruited. MLPA was performed using the P034/P035 DMD (MRC Holland) kit. Multiplex PCR was performed using Chamberlain and Beggs primer sets with appropriate controls. Sequencing of the relevant exon of the DMD gene was planned for discordant results between MLPA and multiplex PCR. SNPs within 8bp from ligation sites on MLPA probes and its frequency in the South Asian population was ascertained from appropriate databases. Single exon deletion was present in 166 (20.4%) of the affected individuals. Validation of MLPA results by multiplex PCR was performed in 135 affected individuals for the following exons- 4, 8, 43, 44, 45, 46, 48, 50, 51, 52 and 53. We obtained 100% concordant results for single exon deletions by MLPA and multiplex PCR probably due to zero frequency of alternate alleles for SNPs in the South Asian population. However, confirmation of MLPA-detected single exon deletions by an alternate technique is still essential due to emerging novel single nucleotide polymorphisms.
 
HTML Full Text  Download PDF