Real Time Paper Based Detection of Streptococcus bovis using Chromogenic Substrate in Resource Constrained Environments

Abstract

Healthcare management faces a variety of difficulties in Circumstances with limited resources. Accurate diagnosis is the first step in disease prevention and therapy that works However, the diagnostic tools that are available in the economically advanced world are frequently of little utility in underdeveloped nations including Pakistan. The gram-positive bacterium Streptococcus bovis is an opportunistic pathogen that can cause various infections from superficial skin infections to severe and potentially fatal invasive diseases. Objective: To design a platform in environments with limited resources for the quick detection of Streptococcus bovis. Methods: A paper based analytical device (PAD) has been created with enrichment in sterile PYP broth for 1.5 hours and used for the detection of alkaline phosphatase activity using the chromogenic substrate Para-Nitrophenyl Phosphate (PNPP), the specie could be found in clinical samples. A coagulase test and sample Gram staining was conducted with the test. Qualitative detection was evaluated by visual detection while quantitative analysis is carried out using Image J software. Results: Alkaline Phosphatase ( S. bovis)  reacts with the PNPP substrate (5.7 mM) in the presence of PAD. Concentrations below 4.5x104 cfu mL resulted in the detection of a color change. The micro PADs were incubated at 37 °C for 3–4 hours before reaction. Colored product (yellow) indicated the presence of S.bovis. Conclusion: Within 2 hours including enrichment time the test may identify Streptococcus bovis up to 104 CFUmL-1