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

Amna  Mahmood; Amtul Jamil Sami

doi:10.54393/pjhs.v3i07.344

Authors

Amna Mahmood School of Biochemistry and Biotechnology, University of the Punjab, Pakistan
Amtul Jamil Sami School of Biochemistry and Biotechnology, University of the Punjab, Pakistan

DOI:

https://doi.org/10.54393/pjhs.v3i07.344

Keywords:

Streptococcus bovis, Paper Based Detection, Bacterial Identification, Diagnosis

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

References

Abou Tayoun AN, Burchard PR, Malik I, Scherer A, Tsongalis GJ. Democratizing molecular diagnostics for the developing world. American journal of clinical pathology. 2014 Jan; 141(1): 17-24. doi: 10.1309/AJCPA1L4KPXBJNPG

Martinez AW, Phillips ST, Wiley BJ, Gupta M, Whitesides GM. FLASH: a rapid method for prototyping paper-based microfluidic devices. Lab on a Chip. 2008; 8(12): 2146-50 doi: 10.1039/b811135a

Shanan S, Gumaa SA, Sandström G, Abd H. Significant association of Streptococcus bovis with malignant gastrointestinal diseases. International journal of microbiology. 2011 Oct; 2011.doi: 10.1155/2011/792019

Gómez-Garcés JL, Gil Y, Burillo A, Wilhelmi I, Palomo M. Diseases associated with bloodstream infections caused by the new species included in the old Streptococcus bovis group. Enfermedades Infecciosas y Microbiologia Clinica. 2012 Feb; 30(4): 175-9. doi: 10.1016/j.eimc.2011.09.015

Spellerberg B and Brandt C. Laboratory diagnosis of Streptococcus pyogenes (group A streptococci). Streptococcus pyogenes: Basic Biology to Clinical Manifestations. 2022 Oct. 2nd edition

Zhang C, Mcadams DA, Grunlan JC. Nano/micro‐manufacturing of bioinspired materials: a review of methods to mimic natural structures. Advanced Materials. 2016 Aug; 28(30): 6292-321. doi: 10.1002/adma.201604494

Gurunathan S, Park JH, Han JW, Kim JH. Comparative assessment of the apoptotic potential of silver nanoparticles synthesized by Bacillus tequilensis and Calocybe indica in MDA-MB-231 human breast cancer cells: targeting p53 for anticancer therapy. International journal of nanomedicine. 2015; 10: 4203 doi: 10.2147/IJN.S83953

Lin PC, Lin S, Wang PC, Sridhar R. Techniques for physicochemical characterization of nanomaterials. Biotechnology advances. 2014 Jul; 32(4): 711-26. doi: 10.1016/j.biotechadv.2013.11.006

Kalimuthu K, Babu RS, Venkataraman D, Bilal M, Gurunathan S. Biosynthesis of silver nanocrystals by Bacillus licheniformis. Colloids and surfaces B: Biointerfaces. 2008 Aug 1; 65(1): 150-3. doi: 10.1016/j.colsurfb.2008.02.018.

Nair PM and Chung IM. Physiological and molecular level effects of silver nanoparticles exposure in rice (Oryza sativa L.) seedlings. Chemosphere. 2014 Oct; 112: 105-13. doi: 10.1016/j.chemosphere.2014.03.056

Martinez AW, Phillips ST, Whitesides GM, Carrilho E. Diagnostics for the Developing World: Microfluidic Paper-Based Analytical Devices. Analytical Chemistry. 2010. 82(1), 3-10. doi: 10.1021/ac9013989

Mackey TK, Liang BA, Cuomo R, Hafen R, Brouwer KC, Lee DE. Emerging and reemerging neglected tropical diseases: a review of key characteristics, risk factors, and the policy and innovation environment. Clinical microbiology reviews. 2014 Oct; 27(4): 949-79. doi: 10.1128/CMR.00045-14

Kamel MI. A view of the health services after COVID-19: an Egyptian perspective. Alexandria Journal of Medicine. 2020 Jul; 56(1): 118-29. doi: 10.1080/20905068.2020.1789391

Laxminarayan R, Duse A, Wattal C, Zaidi AK, Wertheim HF, Sumpradit N, Vlieghe E, Hara GL, Gould IM, Goossens H, Greko C. Antibiotic resistance—the need for global solutions. The Lancet infectious diseases. 2013 Dec; 13(12): 1057-98. doi: 10.1016/S1473-3099(13)70318-9

Boleij A, van Gelder MM, Swinkels DW, Tjalsma H. Clinical Importance of Streptococcus gallolyticus infection among colorectal cancer patients: systematic review and meta-analysis. Clinical Infectious Diseases. 2011 Nov; 53(9): 870-8. doi: 10.1093/cid/cir609

Kawai S, Suzuki H, Yamamoto K, Inui M, Yukawa H, Kumagai H. Purification and characterization of a malic enzyme from the ruminal bacterium Streptococcus bovis ATCC 15352 and cloning and sequencing of its gene. Applied and Environmental Microbiology. 1996 Aug; 62(8): 2692-700. doi: 10.1128/aem.62.8.2692-2700.1996

Lokur A and Ravindra P. Rapid detection of staphylococcus aureus using paper based microfluidic devices for resource limited settings. International Journal of Advanced Research. 2016; 4(10): 818-825.

Galia L, Ligozzi M, Bertoncelli A, Mazzariol A. Real-time PCR assay for detection of Staphylococcus aureus, Panton-Valentine Leucocidin and Methicillin Resistance directly from clinical samples. AIMS microbiology. 2019; 5(2): 138-146. doi: 10.3934/microbiol.2019.2.138

Song B, Wang J, Yan Z, Liu Z, Pan X, Zhang Y, Zhang X. Microfluidics for the rapid detection of Staphylococcus aureus using antibody-coated microspheres. Bioengineered. 2020 Jan 1; 11(1): 1137-45 doi: 10.1080/21655979.2020.1831362

Noiphung J and Laiwattanapaisal W. Multifunctional paper-based analytical device for in situ cultivation and screening of Escherichia coli infections. Scientific reports. 2019 Feb; 9(1): 1-0. doi: 10.1038/s41598-018-38159-1