Rapid and sensitive detection of Escherichia coli O157:H7 using coaxial channel-based DNA extraction and microfluidic PCR

Four commercial DNA extraction methods, PrepMan Ultra (Applied Biosystems), InstaGene Matrix (BioRad), DNeasy Tissue kit (Qiagen), and UltraClean (MoBio), were tested for PCR detection of Listeria monocytogenes, Escherichia coli O157: H7, Salmonella, and Staphylococcus aureus in fresh, minimally processed vegetables. For comparative purposes, sensitivity assays with specific PCRs were carried out after DNA extraction with the four methods in green pepper, broccoli, and onion artificially inoculated with the four pathogens separately. As confirmed by statistical analysis, the DNeasy Tissue kit rendered the highest sensitivity values in the three matrices assayed for Salmonella, L. monocytogenes, and E. coli O157:H7 and in onion for S. aureus. Despite being the most expensive of the methods compared, the DNeasy Tissue Kit can be successfully applied for any of the four most commonly studied pathogens, thus saving time and overall reducing the cost of the analysis.

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A comparison of DNA extraction and purification methods to detect Escherichia coli O157:H7 in cattle manure

Journal of Microbiological Methods ◽

10.1016/s0167-7012(03)00017-4 ◽

2003 ◽

Vol 54 (2) ◽

pp. 165-175 ◽

Cited By ~ 21

Author(s):

Teegan Trochimchuk ◽

John Fotheringham ◽

Edward Topp ◽

Heidi Schraft ◽

Kam Tin Leung

Keyword(s):

Escherichia Coli ◽

Dna Extraction ◽

Cattle Manure ◽

Escherichia Coli O157 ◽

Extraction And Purification ◽

Purification Methods

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Prevalence of Escherichia coli O157 in Cattle and Swine in Central Mexico†

Journal of Food Protection ◽

10.4315/0362-028x-67.10.2274 ◽

2004 ◽

Vol 67 (10) ◽

pp. 2274-2276 ◽

Cited By ~ 28

Author(s):

T. R. CALLAWAY ◽

R. C. ANDERSON ◽

G. TELLEZ ◽

C. ROSARIO ◽

G. M. NAVA ◽

...

Keyword(s):

Escherichia Coli ◽

Limit Of Detection ◽

The United States ◽

Sensitive Detection ◽

Central Mexico ◽

Escherichia Coli O157 ◽

Immunomagnetic Bead ◽

E Coli ◽

Detection Techniques ◽

Swine Farms

Escherichia coli O157:H7 is a foodborne pathogenic bacterium that can reside undetected in the gastrointestinal tract of cattle because colonization by this bacterium is asymptomatic. Recent research has indicated that swine can carry and transmit this pathogen as well. The development of more advanced and sensitive detection techniques has improved the limit of detection and increased sensitivity for this important pathogen. This study was undertaken to determine the prevalence of E. coli O157 in cattle and swine in Mexico with the more sensitive detection technique of immunomagnetic bead separation. Samples (n = 60 per farm) were taken from four cattle and four swine farms (n = 240 cattle samples, n = 240 swine samples) located throughout central Mexico in October 2001. The prevalence of E. coli O157 was found to be only 1.25% on cattle farms and 2.1% on swine farms. The prevalence in cattle in this study is lower than that reported in the United States and could be related to the lower reported prevalence of E. coli O157 in humans in Mexico. However, further research is needed to verify prevalence throughout other regions of Mexico, as well as prevalence during other seasons of the year.

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A capacitive DNA sensor for sensitive detection of Escherichia coli O157:H7 in potable water based on the z3276 genetic marker: fabrication and analytical performance

The Analyst ◽

10.1039/c9an02291k ◽

2020 ◽

Vol 145 (6) ◽

pp. 2267-2278 ◽

Cited By ~ 4

Author(s):

Rehan Deshmukh ◽

Arun Kumar Prusty ◽

Utpal Roy ◽

Sunil Bhand

Keyword(s):

Escherichia Coli ◽

Genetic Marker ◽

Potable Water ◽

Analytical Performance ◽

Sensitive Detection ◽

Escherichia Coli O157 ◽

Dna Sensor ◽

Label Free ◽

Dna Sensing ◽

Water Based

We report a label-free biosensor for the detection of Escherichia coli O157:H7 ATCC 43895 in potable water using a newly designed DNA sensing probe targeting the z3276 genetic marker.

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