Triplex PCR‐based detection of enterotoxigenic Bacillus cereus ATCC 14579 in nonfat dry milk

Abstract

Although many strains of Bacillaceae are considered nonpathogenic, Bacillus cereus is recognized worldwide as a bacterial pathogen in a variety of foods. The ability of B. cereus to cause gastroenteritis following ingestion of contaminated food is due to the production of enterotoxins. The ubiquity of this genus makes it a persistent problem for quality assurance in food processing environments. The primary objective of this study was to develop and apply a multiplex real-time PCR-based assay for rapid and sensitive detection of enterotoxigenic B. cereus. Template DNA was separately extracted from tryptic soy broth (TSB)-grown and 2.5% Nonfat Dry Milk (NFDM)-grown B. cereus using a commercial system. Three enterotoxin gene fragments (hblC, nheA, and hblA) were simultaneously amplified in real-time followed by melting curve analysis to confirm amplicon identity. Resolution of melting curves (characteristic T(m)) was achieved for each amplicon (hblC = 74.5 °C; nheA = 78 °C; and hblA = 85.5 °C in TSB and 84 °C in NFDM) with an assay sensitivities of 10(1) CFU/ml for both TSB and NFDM-grown B. cereus compared to 10(4) CFU/ml in either matrix using gel electrophoresis. The results demonstrate the potential sensitivity of real-time bacterial detection methods in a heterogenous food matrix using real-time PCR.

Real-Time PCR Detection of Paenibacillus spp. in Raw Milk To Predict Shelf Life Performance of Pasteurized Fluid Milk Products

“…The absence of appropriate tools may reflect, in part, the fact that the bacterial ecology of pasteurized fluid milk has only recently been characterized at the molecular level, which led to identification of Paenibacillus as the predominant fluid milk spore-forming spoilage genera (12,19,21,34). Rapid methods to detect spore-forming bacteria have primarily focused on food-borne pathogens, e.g., Bacillus cereus (13,27,43), that pose a significant health threat. However, the presence of spore-forming bacteria that can resist multiple processing hurdles and affect food product quality represents considerable economic and food security concerns.…”

Prevalence and risk factors for Bacillus cereus in raw milk in Inner Mongolia, Northern China

“…Bacillus cereus is a ubiquitous Gram‐positive spore‐forming bacteria, commonly found in raw or processed foods (Gracias and McKillip ; Berthold‐Pluta et al . ).…”

“…Bacillus cereus is a ubiquitous Gram-positive spore-forming bacteria, commonly found in raw or processed foods (Gracias and McKillip 2011;Berthold-Pluta et al 2015). Bacillus cereus may cause two different types of foodborne disease syndromes: emetic and diarrhoeal (Merzougui et al 2014).…”

Deciphering Microorganisms through Intelligent Image Recognition: Machine Learning and Deep Learning Approaches, Challenges, and Advancements

“…Microorganisms are a diverse group of living organisms that play an integral role in various fields such as healthcare, environmental monitoring, and biotechnology [1].The accurate identification and classification of microorganisms are critical for different applications, including clinical microbiology, agriculture, and food production [2].The advantages of microorganisms are many, including their ability to produce useful products like antibiotics, enzymes, and biofuels through biotechnology [3].Microorganisms can also be employed in environmental monitoring, waste management, and food production [4].In agriculture, microorganisms play a vital role in enhancing soil quality, promoting plant growth, and protecting crops from pests and diseases [5].Therefore, it is essential to accurately identify and classify microorganisms for their diverse applications. Microorganisms have numerous benefits and play a crucial role in several fields, including biotechnology, environmental monitoring, and agriculture.…”