Non-invasive genetic diagnosis of Duchenne Muscular Dystrophy probands using salivary DNA

Abstract

Duchenne Muscular Dystrophy (DMD) is the most common fatal X-linked recessive disorder, which means that the disease is carried by mother but usually appears in boys. Since muscle wasting occurs in these children, it is difficult to draw blood from them for the DNA test and also it is an invasive technique. The present study is to bring out the non-invasive method of DNA test for the children. Saliva also harbors a wide spectrum of genetic data that can be used for genetic research and clinical diagnostic applications. Saliva samples from several DMD patients and non DMD patients (as control) were collected and the total DNA was extracted by the modified method and their quality were also determined by spectroscopic method. The resultant DNA was of better quality and good for amplification of the DMD exons by PCR, both uniplex and multiplex. But only bands of less intensity were seen on the gel which may be due to PCR inhibitors and this was further confirmed by PCR method. DNA isolated by this method does not have PCR inhibitors and this was shown by the fact that both multiplex and uniplex PCR’s showed a significant level of amplicons. This study has shown that saliva could be used as a source of genomic DNA for use in PCR based genetic analysis. If standardized, this could be cost effective and less invasive and will benefit the children and old patient too for the diagnosis of all genetic disorders. Thus, this study may be used for the analysis of disease diagnosis and genetic disorder using saliva DNA.