International Journal of Multidisciplinary Health Sciences and Research

Category: Original Article,      DOI: Recived: 19/08/2024, Accepted: 24/10/2024, Published online: 29/10/2024

DNA extraction from bacterial cells using human saliva as a lysis agent

Abhishek Kumar Mishra1*, Dharmsheel Shrivastav2, Kirti Pundir3

Abstract:

Deoxyribonucleic acid (DNA) is the starting biomolecule of central dogma and plays a crucial role in the diagnostic world. Amplification of a specific region of DNA for pathogens is the key to diagnosis. Several manuals, as well as kit-based methods, are available for DNA extraction. Manual methods require a lot of chemicals and enzymes. They are exhaustive, time taking and prone to laboratory accidents (Phenol based DNA extraction). Kit based methods are rapid and easy to perform but are very expensive for low budget laboratories. Under these circumstances, there is a need to establish a quick and cost-effective DNA extraction method in routine. Human saliva is a rich source of several proteins, enzymes (Lysozymes), minerals and electrolytes and has been used as a noninvasive specimen source for a long time. Saliva protects us from pathogenic bacteria by destroying their membrane/cell wall. Considering the killing action of saliva to microbes, it has been used as a substitute for lysis agents/ buffers required for cell lysis of bacteria in the procedure of DNA extraction from Gram positive and Gram negative bacteria. In the saliva method, the time and cost of DNA extraction were found less than in the Phenol Chloroform method. In contrast, not much difference was observed in the concentration of extracted DNA using both these methods. In the Polymerase Chain Reaction (PCR) of extracted DNA, specific amplified bands were seen for each bacterium. Therefore, human saliva may be used as a replacement of lysis buffer for DNA extraction from bacteria