Prevalence and Toxigenic Profiles of Bacillus cereus Isolated from Dried Red Peppers, Rice, and Sunsik in Korea

2009 ◽  
Vol 72 (3) ◽  
pp. 578-582 ◽  
Author(s):  
SUNG KI KIM ◽  
KWANG-PYO KIM ◽  
SUNG SIK JANG ◽  
EUN MI SHIN ◽  
MIN-JEONG KIM ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Multiplex Pcr ◽  
Foodborne Pathogen ◽  
Food Poisoning ◽  
Pcr Assay ◽  
Heat Stable ◽  
Multiplex Pcr Assay ◽  
Red Peppers ◽  
Short Time

Bacillus cereus is a spore-forming foodborne pathogen responsible for diarrheal and emetic types of food poisoning. Intoxication is caused by various enterotoxins or by emetic toxin. Because of its widespread presence and the ability to form heat-stable endospores in a relatively short time, B. cereus has been difficult to control. In this study, 21 rice and 36 Sunsik (a mixture of powdered raw grains) samples were examined for the prevalence of B. cereus. A multiplex PCR assay was used to evaluate the distribution of 10 different toxigenicity-related genes among 1,082 B. cereus strains isolated from dried red peppers (919 isolates), rice (98 isolates), and Sunsik (65 isolates). The results suggest that (i) the examined foods were free of the emetic toxin but not free of enterotoxins and (ii) the distribution of enterotoxigenic genes was significantly different among the B. cereus isolates from various sources.

Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

2008 ◽  
Vol 71 (10) ◽  
pp. 2094-2099 ◽  
Author(s):  
YU-CHANG CHANG ◽  
JAN-YI WANG ◽  
AMMAIYAPPAN SELVAM ◽  
SHU-CHEN KAO ◽  
SHANG-SHYNG YANG ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diarrheal Disease ◽  
Simultaneous Detection ◽  
Food Poisoning ◽  
Food Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Causative Agents ◽  
Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

Improved multiplex PCR assay for simultaneous detection of Bacillus cereus emetic and enterotoxic strains

2012 ◽  
Vol 21 (5) ◽  
pp. 1439-1444 ◽  
Author(s):  
Jae-Myung Kim ◽  
Fereidoun Forghani ◽  
Jung-Beom Kim ◽  
Yong-Bae Park ◽  
Myoung-Su Park ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Establishment of a Novel Multiplex PCR Assay and Detection of Toxigenic Strains of the Species in the Bacillus cereus Group

2005 ◽  
Vol 68 (10) ◽  
pp. 2123-2130 ◽  
Author(s):  
I-CHEN YANG ◽  
DANIEL YANG-CHIH SHIH ◽  
TSUI-PING HUANG ◽  
YUN-PU HUANG ◽  
JAN-YI WANG ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Multiplex Pcr ◽  
Food Poisoning ◽  
Assay Method ◽  
Toxin Gene ◽  
Taq Polymerase ◽  
Toxin Genes ◽  
Multiplex Pcr Assay ◽  
Group A ◽  
Toxigenic Strains

Five different enterotoxins and one emetic toxin of Bacillus cereus have been characterized. To amplify all of the enterotoxin and emetic-specific sequences of the species in the B. cereus group, a multiplex PCR with 12 primer pairs was established. In developing the assay method, a common terminal sequence at the 3′ ends of all primers was chosen and a hot start Taq polymerase was used to overcome primer dimer formation. The assay was successfully applied to analyze the toxigenic potential of 162 food-poisoning and food-related strains. Results showed that there were 10 toxigenic patterns for all the test strains. All of the B. cereus strains carried at least one toxin gene. More than 70% of Bacillus mycoides strains carried no known toxin genes. The toxin profiles and toxin genes of B. mycoides strains were significantly different from B. cereus strains (P < 0.05), although the two species were closely related. The results suggest that many B. mycoides strains might be less prone to cause food poisoning. They also indicate the importance of detecting the toxin genes together with the detection of the species in the B. cereus group.

Prevalence and Genetic Diversity of Bacillus cereus in Dried Red Pepper in Korea

2007 ◽  
Vol 70 (4) ◽  
pp. 917-922 ◽  
Author(s):  
EUIYOUNG CHOO ◽  
SUNG SIK JANG ◽  
KYUMSON KIM ◽  
KWANG-GEUN LEE ◽  
SUNGGI HEU ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Bacillus Cereus ◽  
Multiplex Pcr ◽  
Random Amplified Polymorphic Dna ◽  
Bacterial Pathogen ◽  
Culture Method ◽  
Average Concentration ◽  
Red Pepper ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Bacillus cereus is a foodborne spore-forming bacterial pathogen that is ubiquitous in the natural environment. Infections with this pathogen manifest as diarrheal or emetic types of food poisoning. In this study, 140 samples of dried red pepper purchased in Korea were assayed for the presence of B. cereus according to the U.S. Food and Drug Administration standard culture method. A multiplex PCR assay was developed for the rapid confirmation of B. cereus as an alternative to conventional biochemical confirmation tests. The genetic diversity of B. cereus isolates was investigated using a random amplified polymorphic DNA (RAPD) assay. B. cereus was found in 84.3% of the dried red pepper samples, with an average concentration of 1.9 × 104 CFU/g. B. cereus could be detected and distinguished from B. thuringiensis in the multiplex PCR assay by using the BCFW1 plus BCrevnew and the K3 plus K5 primer sets designed to detect the gyrB gene of B. cereus and B. thuringiensis and the cry gene of B. thuringiensis. A RAPD assay using the OPG 16 and MUP 3 primers was used to successfully distinguish among isolates, thus elucidating the genetic diversity of B. cereus isolates. The discriminating ability of the OPG 16 primer (142 types) was about threefold higher than that of MUP 3 (52 types) in the RAPD assay.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

2021 ◽  
pp. 106198
Author(s):  
Sunarno ◽  
Khariri ◽  
Fauzul Muna ◽  
Kambang Sariadji ◽  
Yuni Rukminiati ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
New Approach ◽  
Multiplex Pcr Assay ◽  
Corynebacterium Sp

A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera)

2010 ◽  
Vol 105 (2) ◽  
pp. 151-155 ◽  
Author(s):  
Mollah Md. Hamiduzzaman ◽  
Ernesto Guzman-Novoa ◽  
Paul H. Goodwin
Keyword(s):  
Apis Mellifera ◽  
Multiplex Pcr ◽  
Honey Bees ◽  
Pcr Assay ◽  
Multiplex Pcr Assay
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