A comparison of ELISA and RPLA for detection of Bacillus cereus diarrhoeal enterotoxin

Day, T.; Tatani, S.R.; Notermans, S.; Bennett, Reginald W.

doi:10.1111/j.1365-2672.1994.tb03037.x

Cited by 48 publications

(20 citation statements)

References 17 publications

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“…Several studies have compared the BCET-RPLA and TECRA-VIA kits for their ability to detect the B. cereus enterotoxin (5,8,19). Buchanan and Schultz reported the positive identification of 8 out of 10 and 9 out of 10 enterotoxigenic B. cereus strains by the BCET-RPLA and TECRA-VIA kits, respectively (5).…”

Section: Discussionmentioning

confidence: 99%

Development of a Fluorogenic Probe-Based PCR Assay for Detection of Bacillus cereus in Nonfat Dry Milk

Kim

Czajka

Batt

2000

Appl Environ Microbiol

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A fluorogenic probe-based PCR assay was developed and evaluated for its utility in detecting Bacillus cereus in nonfat dry milk. Regions of the hemolysin and cereolysin AB genes from an initial group of two B. cereus isolates and two Bacillus thuringiensis isolates were cloned and sequenced. Three single-base differences in two B. cereus strains were identified in the cereolysin AB gene at nucleotides 866, 875, and 1287, while there were no species-consistent differences found in the hemolysin gene. A fluorogenic probe-based PCR assay was developed which utilizes the 5-to-3 exonuclease of Taq polymerase, and two fluorogenic probes were evaluated. One fluorogenic probe (cerTAQ-1) was designed to be specific for the nucleotide differences at bases 866 and 875 found in B. cereus. A total of 51 out of 72 B. cereus strains tested positive with the cerTAQ-1 probe, while only 1 out of 5 B. thuringiensis strains tested positive. Sequence analysis of the negative B. cereus strains revealed additional polymorphism found in the cereolysin probe target. A second probe (cerTAQ-2) was designed to account for additional polymorphic sequences found in the cerTAQ-1-negative B. cereus strains. A total of 35 out of 39 B. cereus strains tested positive (including 10 of 14 previously negative strains) with cerTAQ-2, although the assay readout was uniformly lower with this probe than with cerTAQ-1. A PCR assay using cerTAQ-1 was able to detect approximately 58 B. cereus CFU in 1 g of artificially contaminated nonfat dry milk. Forty-three nonfat dry milk samples were tested for the presence of B. cereus with the most-probablenumber technique and the fluorogenic PCR assay. Twelve of the 43 samples were contaminated with B. cereus at levels greater than or equal to 43 CFU/g, and all 12 of these samples tested positive with the fluorogenic PCR assay. Of the remaining 31 samples, 12 were B. cereus negative and 19 were contaminated with B. cereus at levels ranging from 3 to 9 CFU/g. All 31 of these samples were negative in the fluorogenic PCR assay. Although not totally inclusive, the PCR-based assay with cerTAQ-1 is able to specifically detect B. cereus in nonfat dry milk.

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Section: Discussionmentioning

confidence: 99%

Development of a Fluorogenic Probe-Based PCR Assay for Detection of Bacillus cereus in Nonfat Dry Milk

Kim

Czajka

Batt

2000

Appl Environ Microbiol

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“…The two were given parallel evaluations in four studies. 28,[93][94][95] BCET-RPLA (Oxoid) is the most commonly cited test method for diarrhoeal B. cereus, identified in 12 evaluative and non-evaluative studies in the wider search. Fletcher and Logan 95 compared the BCET-RPLA (Oxoid) and BDE-VIA (Bacillus diarrhoeal enterotoxin, Tecra) with an improved McCoy cell culture cytotoxicity assay.…”

Section: Diarrhoeal Strainsmentioning

confidence: 99%

“…They found the cytotoxicity assay was more sensitive than the Oxoid kit and, unlike the Tecra kit, did not give false positive results. Beecher and Wong 93 were similarly unclear as to the usefulness of the two commercial kits, although Day and colleagues 94 suggested that the Tecra kit is more reliable, having detected 87.5% (14/16) strains, compared with the Oxoid kit, which detected only 43.75% (7/16). Another study which examined foods and faeces using Tecra EIA detected the enterotoxin in seven out of 34 foods and 10 out of 15 faecal specimens from outbreaks of food poisoning.…”

Section: Diarrhoeal Strainsmentioning

confidence: 99%

A systematic review of the clinical, public health and cost-effectiveness of rapid diagnostic tests for the detection and identification of bacterial intestinal pathogens in faeces and food

Abubakar¹,

Irvine²,

Aldus³

et al. 2007

Health Technol Assess

103

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“…Traditional methods to detect food-borne bacteria often rely on time-consuming growth in culture media, isolation of bacteria, biochemical identification, and sometimes serology (5,8). There have been many attempts to develop faster, more convenient, more sensitive, and more specific techniques for detection and diagnosis of pathogenic bacteria, including immunological methods, biosensors, and microarray technologies.…”

mentioning

confidence: 99%

Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria

Jin

Yin

2009

Appl Environ Microbiol

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In the present study, a simple and rapid multiplexed bead-based mesofluidic system (BMS) was developed for simultaneous detection of food-borne pathogenic bacteria, including Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii, Shigella, Escherichia coli O157:H7, and Campylobacter jejuni. This system is based on utilization of isothiocyanate-modified microbeads that are 250 m in diameter, which were immobilized with specific amino-modified oligonucleotide probes and placed in polydimethylsiloxane microchannels. PCR products from the pathogens studied were pumped into microchannels to hybridize with the oligonucleotide-modified beads, and hybridization signals were detected using a conventional microarray scanner. The short sequences of nucleic acids (21 bases) and PCR products characteristic of bacterial pathogens could be detected at concentrations of 1 pM and 10 nM, respectively. The detection procedure could be performed in less than 30 min with high sensitivity and specificity. The assay was simple and fast, and the limits of quantification were in the range from 500 to 6,000 CFU/ml for the bacterial species studied. The feasibility of identification of food-borne bacteria was investigated with samples contaminated with bacteria, including milk, egg, and meat samples. The results demonstrated that the BMS method can be used for effective detection of multiple pathogens in different foodstuffs.Bacterial food-borne pathogens pose a significant threat to human and animal heath. These organisms are the leading causes of illness and death in less developed countries, killing approximately 1.8 million people annually, and they are the third leading cause of death. In developed countries, foodborne pathogens are responsible for millions of cases of infectious gastrointestinal diseases each year, costing billions of dollars in medical care and lost productivity (42). The Centers for Disease Control and Prevention estimates that food-borne diseases cause approximately 76 million illnesses, 325,000 hospitalizations, and 5,000 deaths in the United States each year (25). The Food and Drug Administration's 2005 Food Code states that the estimated cost of food-borne illness is $10 billion to $83 billion annually (29). The major food-borne pathogens include Salmonella, Listeria monocytogenes, Staphylococcus aureus, Vibrio parahaemolyticus, Campylobacter jejuni, Escherichia coli O157:H7, Enterobacter sakazakii, and Shigella (14,32,41).Rapid detection and identification of pathogens and other microbial contaminants in food are needed by the food industry, food safety agencies, and public health bureaus. Traditional methods to detect food-borne bacteria often rely on time-consuming growth in culture media, isolation of bacteria, biochemical identification, and sometimes serology (5, 8). There have been many attempts to develop faster, more convenient, more sensitive, and more specific techniques for detection and diagnosis of pathogenic bacteria, including immunological m...

show abstract

A comparison of ELISA and RPLA for detection of Bacillus cereus diarrhoeal enterotoxin

Cited by 48 publications

References 17 publications

Development of a Fluorogenic Probe-Based PCR Assay for Detection of Bacillus cereus in Nonfat Dry Milk

Development of a Fluorogenic Probe-Based PCR Assay for Detection of Bacillus cereus in Nonfat Dry Milk

A systematic review of the clinical, public health and cost-effectiveness of rapid diagnostic tests for the detection and identification of bacterial intestinal pathogens in faeces and food

Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria

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