Development and on-site evaluation of an easy-to-perform and low-cost food pathogen diagnostic workflow for low-resource communities

Abstract

20Food-borne diseases are a leading cause of illness and death in many developing 21 countries and thus, there is a real need to develop affordable and practical 22 technologies that can help improve food safety in these countries. The ability to 23 efficiently identify food pathogens is essential to allow national regulatory authorities 24 to monitor food quality and implement safety protocols. In this study, we have 25 developed a simple, low-cost ($0.76 (USD)) complete food pathogen diagnostic 26 workflow ideally suited for deployment in low-resource environments that uses a 27 simple four step process (sample enrichment, cell lysis, DNA amplification, and 28 naked-eye readout). The minimal number of steps and equipment involved in our 29 diagnostic workflow, as well as the simplicity of the yes/no flocculation readout, 30 allows non-technical personnel to perform and interpret the assay. To evaluate the 31 system's performance, we tested the entire system on fresh produce samples 32 collected from local farms and markets in Cambodia for the presence of the E. coli 33 O157 O-antigen polymerase, wzy. Although this was a proof-of-concept study, our 34 system successfully revealed a clear correlation between the origin and condition of 35 the produce collected and their likelihood of contamination. In conclusion, we believe 36 that our easy-to-perform diagnostic system can have a significant impact on 37 improving food quality and human health if adopted by regulatory authorities in 38 developing countries due to the assay's simplicity, affordability, and adaptability. 3 40 Introduction 41Over the last 20 years, there has been a significant increase in the incidence of food-42 borne diseases worldwide [1]. The risk of infection by a food-borne disease is 43 significantly higher in developing countries due to a combination of factors including 44 access to clean water and bathroom facilities, poor hygiene education, inadequate 45 food production and storage practices, and either insufficient food safety legislation 46 or poor implementation of existing legislation [2]. Most food-borne disease infections 47 in these countries result from the consumption of perishable foods sold in informal 48 markets [3] and, as such, food-borne diseases have become a leading cause of 49 illness and death in developing countries [4, 5]. Hence, there is a need for low-cost 50 and simple technologies that can help health authorities to monitor and enforce 51 adequate levels of food safety. In addition to the health benefits, increased food 52 safety standards would likely have economic benefits as a result of increased 53 demand for fresh produce exports [3]. 54 55 Shiga toxin-producing Escherichia coli (STEC) are a class of E. coli responsible for 56 many food-borne outbreaks and sporadic cases of gastrointestinal illness, with a 57 range of symptoms including haemorrhagic colitis (stomach cramping and bloody 58 diarrhoea) and the potentially fatal hemolytic-uremic syndrome (break down of red 59 blood cells, kidney failure, re...

Investigating and Modeling the Factors That Affect Genetic Circuit Performance

“…Synthetic biology aims to address pressing global challenges including disease diagnosis and treatment, , biofuel production, − contamination detection, and biomanufacturing. ,− These are achieved by engineering biological systems with new capabilities, granting cellular control and user-defined performance. , The variety of genetic circuit functions include a genetic toggle switch, genetic counters, low- or high-frequency filters, , adders, sequential asynchronous logic circuits, and more.…”

“…Synthetic biology aims to address pressing global challenges including disease diagnosis and treatment, 1,2 biofuel production, 3−5 contamination detection, 6 and biomanufacturing. 4,7−11 These are achieved by engineering biological systems with new capabilities, granting cellular control and user-defined performance.…”

Investigating and Modeling the Factors that Affect Genetic Circuit Performance

“…Synthetic biology aims to address pressing global challenges including disease diagnosis and treatment [1, 2], bio-fuels production [3, 4, 5], contamination detection [6], bio-manufacturing [7, 8, 9, 4, 10, 11], etc. These are achieved by engineering biological systems with new capabilities, granting cellular control and user-defined performance [12, 4].…”