Abstract
Clostridium perfringens α toxin (CPA) is an important virulence factor that causes livestock hemorrhagic enteritis and food poisoning by contaminated meat products. In this study, the nano-silica microspheres combined with smartphone image processing technology was developed to realize real-time CPA detection. First, the N-terminal and C-terminal domain of the CPA toxin (CPAC3 and CPAN) and their anti-sera were prepared. The silica microspheres coupled with the antibody of CPAC3 was prepared to capture the toxin that existed in the detection sample and the fluorescent-labeled antibody of CPAN was incubated. Moreover, the fluorescent pictures of gray value were performed in a cell phone app, corresponding to toxin concentration. The new assay takes 90 min to perform and can detect CPA as little as 32.8 ng/mL. Our results showed a sensitive, stable, and convenient CPA detection system, which provides a novel detection method of native CPA in foods.
Highlights
Clostridium perfringens (C. perfringens, named Clostridium welchii) can result in food-borne gastroenteritis and other infectious diseases
The 5 μg/mL purified CPAC3 or CPAN protein diluted with carbonate buffer was added into a 96-well plate at 100 μL/well and stayed overnight at 4 ◦C; the plate was washed with 0.1% Tween-20 PBS buffer three three times, and continued to block it with 1% bovine serum albumin (BSA)-PBST buffer at 37 ◦C for 30 min
N-terminal and C-terminal alone have no hemolytic activity and lethal activity, and only their synergy can have the activity of α toxin itself
Summary
Clostridium perfringens (C. perfringens, named Clostridium welchii) can result in food-borne gastroenteritis and other infectious diseases. As the most important virulence factor of C. perfringens type A [3], α toxin has the characteristics of cytotoxicity, hemolytic activity, lethality, skin necrosis, myonecrosis, granulopoiesis [4], inhibition of erythroid differentiation [5], platelet aggregation, and increased vascular permeability. It can be found in the small intestines of domestic animals and can contaminate many types of retail meat products, milk, and dairy products, leading to food poisoning [6]. Our study establishes a fast, convenient, and visual detection method for foodborne pathogenic CPA, which provides a new idea for immunoassay of other soluble toxins
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