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 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.

scholarly journals Development of RT-multiplex PCR Assay for Detection of Adenovirus and Group A and C Rotaviruses in Diarrheal Fecal Specimens from Children in China

2004 ◽  
Vol 78 (8) ◽  
pp. 699-709 ◽  
Author(s):  
Hainian YAN ◽  
Tuan Anh NGUYEN ◽  
Tung Gia PHAN ◽  
Shoko OKITSU ◽  
Yan LI ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Group A

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.

scholarly journals DETECTION OF HBLD TOXIN GENE BY BACILLUS CEREUS ISOLATED FROM MEAT CURRY FOOD SAMPLES IN MALAYSIAN RESTAURANTS

2020 ◽  
Vol 4 (2) ◽  
pp. 68-72
Author(s):  
Marwan Msarah ◽  
Ahmed Alsier ◽  
Sahilah, A.M.
Keyword(s):  
Bacillus Cereus ◽  
Foodborne Pathogen ◽  
Food Poisoning ◽  
Egg Yolk ◽  
Food Samples ◽  
Blood Agar ◽  
Toxin Gene ◽  
Biochemical Tests ◽  
Isolation Medium ◽  
Hemolysin Bl

Bacillus cereus is a ubiquitous foodborne pathogen, can cause food poisoning, leading to infections, have two major types of food poisoning emetic and diarrheal. Foods rich in protein such as meat are associated with foodborne outbreaks of diarrhea caused by B. cereus. The aim of this study is to isolate and identify B. cereus from ready to eat (RTE) meat curry from restaurants in Malaysia and to detect hblD pathogenic gene of B. cereus isolates. Mannitol egg yolk polymyxin agar was used as a selective isolation medium. Commercially available kits and boiling methods were used for DNA extraction, samples acquired from restaurants were examined for the presence of Hemolysin BL gene by polymerase chain reaction (PCR). Among all isolates, twenty-four of B. cereus isolates detected for HBL enterotoxin production by the discontinuous pattern on HBL sheep blood agar then confirmed by biochemical tests. More than 58.33 % of the isolate showed discontinuous hemolysis pattern on HBl blood agar and 29.16% of the samples were shown positive for hblD gene that can cause diarrhea with the size of 807bp on gel. This study demonstrated that RTE meat curry was a potential source for entero-toxigenic B. cereus and the presence of the hblD toxin genes for the HBL complex in the isolates tested were highly associated. Therefore, these meat curry isolates should be regarded as potential toxin producers.

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

scholarly journals Development of a Single-Reaction Multiplex PCR Toxin Typing Assay for Staphylococcus aureusStrains

2000 ◽  
Vol 66 (4) ◽  
pp. 1347-1353 ◽  
Author(s):  
Naresh K. Sharma ◽  
Catherine E. D. Rees ◽  
Christine E. R. Dodd
Keyword(s):  
Multiplex Pcr ◽  
Toxin Gene ◽  
Gene Products ◽  
Pcr Assay ◽  
Specific Primers ◽  
Purification Method ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Immunological Tests ◽  
Single Reaction

ABSTRACT We describe here the development of a single-reaction multiplex PCR assay for the enterotoxin genes from Staphylococcus aureusthat utilizes a universal toxin gene primer in combination with toxin-specific primers to amplify characteristic toxin gene products. In combination with a new DNA purification method, the assay can detect enterotoxin genes A to E from a pure culture within 3 to 4 h. The test was used to characterize a diverse set of environmental S. aureus isolates, and a 99% correlation with toxin typing using standard immunological tests was found. The design of the assay allows it to be extended to include both newly characterized and as-yet-unknown toxin genes.

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