An Aggregation-Induced Emission Material Labeling Antigen-Based Lateral Flow Immunoassay Strip for Rapid Detection of Escherichia coli O157:H7

2021 ◽  
pp. 247263032098193
Author(s):  
Cheng Liu ◽  
Shuiqin Fang ◽  
Yachen Tian ◽  
Youxue Wu ◽  
Meijiao Wu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Foodborne Pathogen ◽  
Test Strip ◽  
Lateral Flow ◽  
Detection Sensitivity ◽  
Lateral Flow Immunoassay ◽  
Escherichia Coli O157 ◽  
Aggregation Induced Emission ◽  
E Coli

Escherichia coli O157:H7 ( E. coli O157:H7) is a dangerous foodborne pathogen, mainly found in beef, milk, fruits, and their products, causing harm to human health or even death. Therefore, the detection of E. coli O157:H7 in food is particularly important. In this paper, we report a lateral flow immunoassay strip (LFIS) based on aggregation-induced emission (AIE) material labeling antigen as a fluorescent probe for the rapid detection of E. coli O157:H7. The detection sensitivity of the strip is 105 CFU/mL, which is 10 times higher than that of the colloidal gold test strip. This method has good specificity and stability and can be used to detect about 250 CFU of E. coli O157:H7 successfully in 25 g or 25 mL of beef, jelly, and milk. AIE-LFIS might be valuable in monitoring food pathogens for rapid detection.

Functional nanozyme mediated multi-readout and label-free lateral flow immunoassay for rapid detection of Escherichia coli O157:H7

2020 ◽  
Vol 329 ◽  
pp. 127224
Author(s):  
Zonghan Wang ◽  
Xiaolin Yao ◽  
Yongzhi Zhang ◽  
Rong Wang ◽  
Yanwei Ji ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Escherichia Coli O157 ◽  
Label Free

Potential pitfalls in the quantitative molecular detection ofEscherichia coliO157:H7 in environmental matrices

2006 ◽  
Vol 52 (5) ◽  
pp. 482-488 ◽  
Author(s):  
Rebekka R.E Artz ◽  
Lisa M Avery ◽  
Davey L Jones ◽  
Ken Killham
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Environmental Samples ◽  
Detection Sensitivity ◽  
Escherichia Coli O157 ◽  
Soil System ◽  
E Coli ◽  
Animal Wastes ◽  
Pcr Assays

The detection sensitivity and potential interference factors of a commonly used assay based on real-time polymerase chain reaction (PCR) for Escherichia coli O157:H7 using eae gene-specific primers were assessed. Animal wastes and soil samples were spiked with known replicate quantities of a nontoxigenic strain of E. coli O157:H7 in a viable or dead state and as unprotected DNA. The detection sensitivity and accuracy of real-time PCR for E. coli O157:H7 in animal wastes and soil is low compared to enrichment culturing. Nonviable cells and unprotected DNA were shown to produce positive results in several of the environmental samples tested, leading to potential overestimates of cell numbers due to prolonged detection of nonviable cells. This demonstrates the necessity for the specific calibration of real-time PCR assays in environmental samples. The accuracy of the eae gene–based detection method was further evaluated over time in a soil system against an activity measurement, using the bioluminescent properties of an E. coli O157:H7 Tn5luxCDABE construct. The detection of significant numbers of viable but nonculturable (VBNC) as well as nonviable and possibly physically protected cells as shown over a period of 90 days further complicates the use of real-time PCR assays for quick diagnostics in environmental samples and infers that enrichment culturing is still required for the final verification of samples found positive by real-time PCR methods.Key words: Escherichia coli O157:H7, real-time PCR, animal waste, soil, VBNC.

A Comparison of the Survival in Feces and Water of Escherichia coli O157:H7 Grown under Laboratory Conditions or Obtained from Cattle Feces

2006 ◽  
Vol 69 (1) ◽  
pp. 6-11 ◽  
Author(s):  
L. SCOTT ◽  
P. McGEE ◽  
J. J. SHERIDAN ◽  
B. EARLEY ◽  
N. LEONARD
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogen ◽  
Hemorrhagic Colitis ◽  
Contaminated Water ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Cattle Feces ◽  
Oral Inoculation ◽  
Before And After ◽  
Uremic Syndrome

Escherichia coli O157:H7 is an important foodborne pathogen that can cause hemorrhagic colitis and hemolytic uremic syndrome. Cattle feces and fecally contaminated water are important in the transmission of this organism on the farm. In this study, the survival of E. coli O157:H7 in feces and water was compared following passage through the animal digestive tract or preparation in the laboratory. Feces were collected from steers before and after oral inoculation with a marked strain of E. coli O157:H7. Fecal samples collected before cattle inoculation were subsequently inoculated with the marked strain of E. coli O157:H7 prepared in the laboratory. Subsamples were taken from both animal and laboratory-inoculated feces to inoculate 5-liter volumes of water. E. coli O157:H7 in feces survived up to 97 days, and survival was not affected by the method used to prepare the inoculating strain. E. coli O157:H7 survived up to 109 days in water, and the bacteria collected from inoculated cattle were detected up to 10 weeks longer than the laboratory-prepared culture. This study suggests that pathogen survival in low-nutrient conditions may be enhanced by passage through the gastrointestinal tract.

Primers with 5′ Flaps Improve the Efficiency and Sensitivity of Multiplex PCR Assays for the Detection of Salmonella and Escherichia coli O157:H7

2013 ◽  
Vol 76 (4) ◽  
pp. 668-673 ◽  
Author(s):  
CHRIS TIMMONS ◽  
SHEFALI DOBHAL ◽  
JACQUELINE FLETCHER ◽  
LI MARIA MA
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Fold Increase ◽  
Detection Sensitivity ◽  
Escherichia Coli O157 ◽  
Bacterial Cells ◽  
Robust Detection ◽  
E Coli ◽  
Pcr Multiplex ◽  
Pcr Assays

Foodborne illnesses caused by Salmonella enterica and Escherichia coli O157:H7 are worldwide health concerns. Rapid, sensitive, and robust detection of these pathogens in foods and in clinical and environmental samples is essential for routine food quality testing, effective surveillance, and outbreak investigations. The aim of this study was to evaluate the effect on PCR sensitivity of adding a short, AT-rich overhanging nucleotide sequence (flap) to the 5′ end of PCR primers specific for the detection of Salmonella and E. coli O157:H7. Primers targeting the invA gene of Salmonella and the rfbE gene of E. coli O157:H7 were synthesized with or without a 12-bp, AT-rich 5′ flap (5′-AATAAATCATAA-3′). Singleplex PCR, multiplex PCR, and real-time PCR sensitivity assays were conducted using purified bacterial genomic DNA and crude cell lysates of bacterial cells. The effect of background flora on detection was evaluated by spiking tomato and jalapeno pepper surface washes with E. coli O157:H7 and Salmonella Saintpaul. When targeting individual pathogens, end-point PCR assays using flap-amended primers were more efficient than nonamended primers, with 20.4 and 23.5% increases in amplicon yield for Salmonella and E. coli O157:H7, respectively. In multiplex PCR assays, a 10- to 100-fold increase in detection sensitivity was observed when the primer flap sequence was incorporated. This improvement in both singleplex and multiplex PCR efficiency and sensitivity can lead to improved Salmonella and E. coli O157:H7 detection.

scholarly journals Rapid Detection of Escherichia coli O157 and Shiga Toxins by Lateral Flow Immunoassays

Toxins ◽  
2016 ◽  
Vol 8 (4) ◽  
pp. 92 ◽  
Author(s):  
Jinliang Wang ◽  
Robab Katani ◽  
Lingling Li ◽  
Narasimha Hegde ◽  
Elisabeth Roberts ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Lateral Flow ◽  
Escherichia Coli O157 ◽  
Shiga Toxins

A remarkable sensitivity enhancement in a gold nanoparticle-based lateral flow immunoassay for the detection of Escherichia coli O157:H7

RSC Advances ◽  
2015 ◽  
Vol 5 (56) ◽  
pp. 45092-45097 ◽  
Author(s):  
Xi Cui ◽  
Youju Huang ◽  
Jingyun Wang ◽  
Lei Zhang ◽  
Yun Rong ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gold Nanoparticle ◽  
Quantitative Data ◽  
Visual Inspection ◽  
Sensitivity Enhancement ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Escherichia Coli O157

The size and uniformity of AuNPs were optimized to maximally amplify both visual inspection signals and quantitative data of LFA.

Optimization of Rapid Detection of Escherichia coli O157:H7 and Listeria monocytogenes by PCR and Application to Field Test

2004 ◽  
Vol 67 (8) ◽  
pp. 1634-1640 ◽  
Author(s):  
GI-SEONG MOON ◽  
WANG JUNE KIM ◽  
WEON-SUN SHIN
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Field Test ◽  
Rapid Detection ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Field Samples ◽  
Standard Culture ◽  
Pcr Method ◽  
Primer Sets

For rapid detection of Escherichia coli O157:H7 and Listeria monocytogenes, simple methods for sample preparation and PCR were established and applied to a field test. To improve specificity, primer sets LP43-LP44 and C(+)-D(−) were selected for E. coli O157:H7 and L. monocytogenes, respectively. Through centrifugation and partial heat treatment after enrichment,E. coli O157:H7 and L. monocytogenes were detected at 1 initial CFU without genomic DNA extraction in the culture and with artificially inoculated food samples including milk, chicken, ham, and pork. Based on the optimized PCR method, a feasibility test was carried out using randomly collected field samples. To remove false positives and false negatives, a PCR method using several primer sets, including the optimized primer set, and a standard culture method were used. With the PCR detection and standard culture methods, two pork samples were positive for L. monocytogenes after enrichment, indications that the PCR assay could be effectively used for rapid, sensitive, and species-specific detection of foodborne pathogens.

Incidence and Toxin Production Ability of Escherichia coli O157:H7 Isolated from Cattle Trucks†

2007 ◽  
Vol 70 (10) ◽  
pp. 2383-2385 ◽  
Author(s):  
E. P. CUESTA ALONSO ◽  
S. E. GILLILAND ◽  
C. R. KREHBIEL
Keyword(s):  
Escherichia Coli ◽  
Potential Risk ◽  
Foodborne Pathogen ◽  
Toxin Production ◽  
Escherichia Coli O157 ◽  
Foodborne Illnesses ◽  
E Coli

Twelve cattle trucks were analyzed for the presence of Escherichia coli O157:H7. Three of them had been washed prior to arrival, and the others had not. Seventy-five percent of the trailers were positive for the presence of this foodborne pathogen. A total of 54 cultures were isolated and identified as E. coli O157:H7, all from the trucks that had not been cleaned. Most of the cultures (96.4%) produced Shiga-like toxin (verotoxin). No E. coli O157:H7 was detected in cattle trucks that were cleaned before arrival at the cattle pens. The incidence of E. coli O157:H7 in transport trailers increases the potential risk of contamination of cattle and transmission from farms to feedlots and to packing plants. This contamination increases the potential of contamination of meat during harvest and the risk of foodborne illnesses.

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