Study of Real-Time PCR Assays for Rapid Detection of Food-Borne Pathogens

Fukushima, Hiroshi; Tsunomori, Yoshie

doi:10.11150/kansenshogakuzasshi1970.79.644

Cited by 13 publications

(15 citation statements)

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“…Salmonella, one of the most common causes of food-borne disease outbreaks due to its widespread occurrence and several sources have been known to harbor this pathogen [98]. A duplex real-time SYBR Green LightCycler PCR (LC-PCR) assay was developed for 17 food/water borne bacterial pathogens from stools by Fukushima and co-workers [99][100] [101]. Similarly, RT PCR has been used to quantify the food-borne pathogen Listeria monocytogenes by first incorporating an IAC [102].…”

Section: Food Microbiology and Safetymentioning

confidence: 99%

Real-Time PCR: Revolutionizing Detection and Expression Analysis of Genes

et al. 2007

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Invention of polymerase chain reaction (PCR) technology by Kary Mullis in 1984 gave birth to real-time PCR. Real-time PCR - detection and expression analysis of gene(s) in real-time - has revolutionized the 21(st) century biological science due to its tremendous application in quantitative genotyping, genetic variation of inter and intra organisms, early diagnosis of disease, forensic, to name a few. We comprehensively review various aspects of real-time PCR, including technological refinement and application in all scientific fields ranging from medical to environmental issues, and to plant.

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Section: Food Microbiology and Safetymentioning

confidence: 99%

Real-Time PCR: Revolutionizing Detection and Expression Analysis of Genes

et al. 2007

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“…A range of PCR assays targeting different genomic fragments have been developed for the detection of P. shigelloides (Table 9) (42,51,221,222,223,224,225). One of the earliest assays was configured by González-Rey et al (42) and targeted a specific variable region of the 23S rRNA gene previously identified by Van Camp et al (226).…”

Section: Laboratory Identification Isolation and Identificationmentioning

confidence: 99%

“…This is an advance on 16S rRNA sequencing, where P. shigelloides previously showed a closer affinity with species of the Enterobacteriaceae than with the Aeromonadaceae (19). The gyrB gene has also been used as a target in a PCR assay to detect P. shigelloides in investigations of foodborne outbreaks (223,224).…”

Section: Laboratory Identification Isolation and Identificationmentioning

confidence: 99%

Plesiomonas shigelloides Revisited

Janda¹,

2016

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SUMMARYAfter many years in the familyVibrionaceae, the genusPlesiomonas, represented by a single species,P. shigelloides, currently resides in the familyEnterobacteriaceae, although its most appropriate phylogenetic position may yet to be determined. Common environmental reservoirs for plesiomonads include freshwater ecosystems and estuaries and inhabitants of these aquatic environs. Long suspected as being an etiologic agent of bacterial gastroenteritis, convincing evidence supporting this conclusion has accumulated over the past 2 decades in the form of a series of foodborne outbreaks solely or partially attributable toP. shigelloides. The prevalence ofP. shigelloidesenteritis varies considerably, with higher rates reported from Southeast Asia and Africa and lower numbers from North America and Europe. Reasons for these differences may include hygiene conditions, dietary habits, regional occupations, or other unknown factors. Other human illnesses caused byP. shigelloidesinclude septicemia and central nervous system disease, eye infections, and a variety of miscellaneous ailments. For years, recognizable virulence factors potentially associated withP. shigelloidespathogenicity were lacking; however, several good candidates now have been reported, including a cytotoxic hemolysin, iron acquisition systems, and lipopolysaccharide. WhileP. shigelloidesis easy to identify biochemically, it is often overlooked in stool samples due to its smaller colony size or relatively low prevalence in gastrointestinal samples. However, one FDA-approved PCR-based culture-independent diagnostic test system to detect multiple enteropathogens (FilmArray) includesP. shigelloideson its panel. Plesiomonads produce β-lactamases but are typically susceptible to many first-line antimicrobial agents, including quinolones and carbapenems.

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“…The published concentrations of foodborne pathogenic bacteria in feces are ＞10 6 CFU/g (＞10 4 CFU/ 40 mg harvested by lightly wiping feces with swabs) in the acute phase of foodborne illness (20). However, fecal samples are not always collected in the acute phase.…”

Section: Resultsmentioning

confidence: 99%

A Rapid and Simple Real-Time PCR Assay for Detecting Foodborne Pathogenic Bacteria in Human Feces

Hanabara

Ueda

2016

Jpn J Infect Dis

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SUMMARY: A rapid, simple method for detecting foodborne pathogenic bacteria in human feces is greatly needed. Here, we examined the efficacy of a method that employs a combination of a commercial PCR master mix, which is insensitive to PCR inhibitors, and a DNA extraction method which used sodium dodecyl benzene sulfonate (SDBS), and Tween 20 to counteract the inhibitory effects of SDBS on the PCR assay. This method could detect the target genes (stx1 and stx2 of enterohemorrhagic Escherichia coli, invA of Salmonella Enteritidis, tdh of Vibrio parahaemolyticus, gyrA of Campylobacter jejuni, ceuE of Campylobacter coli, SEA of Staphylococcus aureus, ces of Bacillus cereus, and cpe of Clostridium perfringens) in a fecal suspension containing 1.0 × 10 1 to 1.0 × 10 3 CFU/ml. Furthermore, the assay was neither inhibited nor influenced by individual differences among the fecal samples of 10 subjects or fecal concentration (40-160 mg/ml in the fecal suspension). When we attempted to detect the genes of pathogenic bacteria in 4 actual clinical cases, we found that this method was more sensitive than standard culture method. These results showed that this assay is a rapid, simple detection method for foodborne pathogenic bacteria in human feces.

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Study of Real-Time PCR Assays for Rapid Detection of Food-Borne Pathogens

Cited by 13 publications

References 0 publications

Real-Time PCR: Revolutionizing Detection and Expression Analysis of Genes

Real-Time PCR: Revolutionizing Detection and Expression Analysis of Genes

Plesiomonas shigelloides Revisited

A Rapid and Simple Real-Time PCR Assay for Detecting Foodborne Pathogenic Bacteria in Human Feces

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