Detection of E. coli O157:H7 and Shigella dysenteriae toxins in clinical samples by PCR-ELISA

Amani, Jafar; Ahmadpour, Askary; Fooladi, Abbas Ali Imani; Nazarian, Shahram

doi:10.1016/j.bjid.2015.02.008

Cited by 45 publications

(32 citation statements)

References 28 publications

(35 reference statements)

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“…It has been reported that S. dysenteriae was detected in feces by immune-magnetic isolation and PCR in about 7 h 5 . As stated by a group, the presence of inhibitors is one of the potential problems of PCR and may cause false results also 1 . Sensitivity of PCR for detection of pathogens in food products depends strongly on the procedure of DNA extraction.…”

Section: Discussionmentioning

confidence: 99%

Detection of Shigella dysenteriae Type 1 in Milk by PCR

Gupta

Dhaked

2017

Def. Life. Sc. Jl.

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Shigellosis is the severest form of bacillary dysentery, a disease limited to humans and certain other primates. Conventional microbiological techniques used for the isolation and biochemical identification of microbes are timeconsuming as well as labour intensive. Due to the fastidious nature of Shigella and the lengthy culture time, a rapid and sensitive system for the detection of bacteria is highly desirable. Nucleic acid-based techniques have enormous potential in the detection of biological weapons because of their specificity, sensitivity, and the speed with which results can be obtained. The application of polymerase chain reaction in sensitive detection of bacterial pathogens in direct food samples is largely affected by the quality of the template DNA. To overcome the shortcomings of conventional methods, simple method of sample preparation that facilitates PCR based detection of Shigella dysenteriae in milk is defined. In present study, gene specific primers were designed to detect stxA gene. The genomic DNA extraction from the spiked milk was carried out using proteinase K, lysozyme, chloroform-isoamyl alcohol and heat treatments. For detection extraction of DNA by addition of chloroform-isoamyl alcohol proved to be the most satisfactory amongst all, with limit of detection of 2.0 x 10 3 cfu/reaction and was completed in 4 h.

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Section: Discussionmentioning

confidence: 99%

Detection of Shigella dysenteriae Type 1 in Milk by PCR

Gupta

Dhaked

2017

Def. Life. Sc. Jl.

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“…Mousavi uated a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O 1 from Iran. In our previous study, PCR-ELISA was used to detect genes encoding shiga toxins1 and 2 from Escherichia coli O157: H7 and other Shiga toxin-producing E. coli (STEC) (33). In our procedure, we used DNA genomic ETEC; we also decreased the number of PCR cycles to five for reducing the reaction time.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Specific Polymerase Chain Reaction-Enzyme Linked Immunosorbent Assay for Detection of Escherichia coli LT Toxin From Clinical Isolates

Esfandiari

Amani

Fouladi

et al. 2016

Arch Clin Infect Dis

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Background: Enterotoxigenic Escherichia coli (ETEC) is the most common agent, which causes diarrhea. ETEC is colonized along the cells and produces enterotoxins leading to diarrhea. Different detection methods have been utilized for detection of ETEC heat Labile Toxin (LT) toxins or respective genes. These methods have disadvantages such as high costs and labor time and limitations in handling many samples simultaneously.

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“…The “gold standard” for detecting E. coli O157:H7 in food samples is a traditional separation-identification method which is time-consuming (at least five days) and laborious (involves pre-enrichment, selective enrichment, culture isolation, and identification). Other methods, including the polymerase chain reaction (PCR) [2,3] and enzyme-linked immunosorbent assay (ELISA) [4,5], also may require laborious procedures and expensive instruments.…”

Section: Introductionmentioning

confidence: 99%

Strategy for Accurate Detection of Escherichia coli O157:H7 in Ground Pork Using a Lateral Flow Immunoassay

Cheng

Chen

Zhang

et al. 2017

Sensors

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Escherichia coli O157:H7 is known to cause serious diseases including hemorrhagic colitis and hemolytic uremic syndrome. A gold nanoparticle lateral flow immunoassay (Au-LFIA) was used to detect Escherichia coli O157:H7 in ground pork samples. False-positive results were detected using Au-LFIA; a Citrobacter freundii strain was isolated from the ground pork samples and identified by using CHROmagarTM plates, API 20E, and 16S RNA sequencing. Since C. freundii showed cross-reactivity with E. coli O157:H7 when Au-LFIA test strips were used, a novel method combining modified enrichment with a lateral flow immunoassay for accurate and convenient detection of E. coli O157:H7 in ground pork was developed in this study to minimize these false positives. MacConkey broth was optimized for E. coli O157:H7 enrichment and C. freundii inhibition by the addition of 5 mg/L potassium tellurite and 0.10 mg/L cefixime. Using the proposed modified enrichment procedure, the false-positive rate of ground pork samples spiked with 100 CFU/g C. freundii decreased to 5%.

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Detection of E. coli O157:H7 and Shigella dysenteriae toxins in clinical samples by PCR-ELISA

Cited by 45 publications

References 28 publications

Detection of Shigella dysenteriae Type 1 in Milk by PCR

Detection of Shigella dysenteriae Type 1 in Milk by PCR

Rapid and Specific Polymerase Chain Reaction-Enzyme Linked Immunosorbent Assay for Detection of Escherichia coli LT Toxin From Clinical Isolates

Strategy for Accurate Detection of Escherichia coli O157:H7 in Ground Pork Using a Lateral Flow Immunoassay

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