Background and objective Salmonella enterica Serovars Paratyphi A, B, and C cause paratyphoid fever, also known as enteric fever, causing symptoms similar to typhoid, such as fevers, chills, and abdominal pain, and can be life-threatening in severe cases. The study utilized nested PCR to efficiently identify small bacterial components, enhancing sensitivity and specificity, saving time, and addressing non-culturable and dead material issues often associated with antibiotic treatment. Material method The study, conducted from 2011 to 2013, was conducted in collaboration with the Departments of Medicine and Paediatrics at Sir Sunder Lal Hospital, Institute of Medical Sciences Banaras Hindu University, Uttar Pradesh. DNA extraction from specimens was performed using phenol-chloroform and proteinase K methods. The Van Zwet et al. method was used to extract DNA from 3 to 5 g of stool, mixed with 10% formal saline and ether, and centrifuged at 3,000 rpm for 5 minutes. The nested PCR product was electrophoresed on a 1.5% agarose gel, analyzed using molecular markers, and visualized under ultraviolet illumination. Result The study involved 130 participants, including 90 clinically suspected typhoid fever cases and 40 healthy controls, of varying ages and sexes. detection of Salmonella paratyphi by nested PCR in blood clots was 25.5%, followed by stool specimens at 42.2% and urine at 42.2%. Conclusion The PCR-based method is more sensitive and provides a fast result for detecting Salmonella paratyphi A in various specimens compared to blood culture, which takes almost a week.