Potential of Local Papaya Latex as an Alternative to Proteinase-K for Genomic DNA Isolation of Klebsiella Pneumoniae Supporting PCR Analysis

Background: Nosocomial infections are a major healthcare problem in hospitals that can increase morbidity, mortality, and healthcare costs. Klebsiella pneumoniae is one of the causative bacteria; therefore, a rapid and accurate detection method such as PCR is required. The success of PCR depends on the quality of extracted DNA. Proteinase-K is commonly used to remove contaminating proteins during DNA isolation; however, its limitations necessitate alternative approaches. Papain derived from raw papaya latex exhibits proteolytic activity and has potential as an alternative to Proteinase-K. Objective: To determine the quality of genomic DNA isolates of K. pneumoniae obtained using Papain enzyme from local papaya latex and to evaluate its performance in PCR amplification. Methods: This study employed a comparative experimental design with qualitative and quantitative approaches. During DNA isolation, Papain was used as the treatment, while Proteinase-K served as the control. DNA quality was assessed based on purity and concentration using a UV-Vis spectrophotometer at wavelengths of 260 nm and 280 nm, and visualized using agarose gel electrophoresis. Results: The mean DNA purity values obtained from the Papain and Proteinase-K treatments were 1.03 ± 0.00 and 1.02 ± 0.01, respectively. Meanwhile, the mean DNA concentrations were 195 ± 0.00 ng/µL and 196.25 ±