Studies have shown that saliva is a potential diagnostic tool for the identification and analysis of several biomarkers in infants, children, adults, and uncooperative patients. With the advent of improved efficiency and accuracy of proteomic, genomic, transcriptome analysis salivary diagnostics turn into a clinical and commercial reality. Advanced molecular technique PCR led to the use of saliva as a source of microbial DNA for the detection of bacteria and viruses. Saliva is increasingly used in DNA analysis as it serves as a useful source of biomarker profiling of oral or systemic diseases, and forensic identification. DNA tests are also a method of detection of HIV infection by recognizing the viral sequences in total salivary DNA amplifying by PCR. Several pathogens and microbiota are also detected by this analysis. The potential advantages of salivary analysis for the diagnosis of systemic disease suggest that further studies are warranted. Recently, new methods have been developed for the collection of saliva-based on the modification of the old expectoration techniques. Noninvasive nature, less hazardous and secure to health care workers, salivary specimen collection for diagnosis of various diseases, and including pandemic coronavirus disease has more advantages and acceptable for patients.