Detection of Viable but Non-Culturable (Vbnc) Escherichia Coli from Beef Liver Using Pma-PCR Assay

2021 ◽  
Vol 69 (2) ◽  
pp. 1
Author(s):  
Diaa Ghit
Keyword(s):  
Escherichia Coli ◽  
Pcr Assay ◽  
Beef Liver

Simple, Rapid, and Reliable Detection of Escherichia coli O26 Using Immunochromatography

2013 ◽  
Vol 76 (5) ◽  
pp. 748-754 ◽  
Author(s):  
TARO YONEKITA ◽  
TATSUYA FUJIMURA ◽  
NAOKI MORISHITA ◽  
TAKASHI MATSUMOTO ◽  
FUMIKI MORIMATSU
Keyword(s):  
Escherichia Coli ◽  
Diarrheal Disease ◽  
Culture Method ◽  
Food Samples ◽  
Pcr Assay ◽  
Beef Liver ◽  
E Coli ◽  
Pure Cultures ◽  
Alfalfa Sprout ◽  
Meat Samples

Shiga toxin–producing Escherichia coli (STEC) O26 has been increasingly associated with diarrheal disease all over the world. We developed an immunochromatographic (IC) strip for the rapid detection of E. coli O26 in food samples. To determine the specificity of the IC strip, pure cultures of 67 E. coli and 22 non–E. coli strains were tested with the IC strip. The IC strip could detect all (18 of 18) E. coli O26 strains tested and did not react with strains of any other E. coli serogroup or non–E. coli strains tested (0 of 71). The minimum detection limits for E. coli O26 were 2.2 ×103 to 1.0 ×105 CFU/ml. To evaluate the ability of the IC strip to detect E. coli O26 in food, 25-g food samples (ground beef, beef liver, ground chicken, alfalfa sprout, radish sprout, spinach, natural cheese, and apple juice) were spiked with E. coli O26. The IC strip was able to detect E. coli O26 at very low levels (approximately 1 CFU/25 g of food samples) after an 18-h enrichment, and the IC strip results were in 100% agreement with the results of the culture method and PCR assay. When 115 meat samples purchased from supermarkets were tested, 5 were positive for E. coli O26 with the IC strip; these results were confirmed with a PCR assay. These results suggest that the IC strip is a useful tool for detecting E. coli O26 in food samples.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

scholarly journals A novel report of colistin-resistant Escherichia coli carrying mcr-1 gene from animal and human feacal samples in Nigeria

2018 ◽  
Vol 1 (1) ◽  
pp. 7-10 ◽  
Author(s):  
Olugbenga A. Olowe ◽  
Rita A. Olowe ◽  
Adeolu S. Oluremi ◽  
Olusolabomi J. Adefioye
Keyword(s):  
Escherichia Coli ◽  
Animal Husbandry ◽  
Multiple Drug ◽  
Colistin Resistance ◽  
Pcr Assay ◽  
Southwestern Nigeria ◽  
E Coli ◽  
Multiple Drug Resistant ◽  
Microbiological Methods ◽  
Faecal Samples

Background: The mobilized colistin resistance (m cr)-1 gene confers transferable colistin resistance. Reports of mcr-1-positive Escherichia coli (MCRPE) have attracted substantial attention. However, in Nigeria, there is no report of mcr-1 gene resistance. Since colistin is a last resort for multiple drug-resistant isolates, this study therefore report the prevalence of mcr-1 gene among E. coli isolated from human and animal sources. Methods: Out of a total of 280 samples collected from animal and hum an faecal samples from selected farms in Oyo and Osun States, Southwestern Nigeria between July 2015 and June 2016, 60 E. coli were identified using standard microbiological methods. The mcr-1 gene was detected in the isolates by conventional PCR assay. Results: The m cr-1 gene was low and not statistically significant (p≥0.05). It was detected in 5 (8.3%) of 60 E. coli isolates (4= animals; 1= human) Conclusion: This study is the first report of mcr -1 gene from E. coli from human and animal sources in Nigeria. This calls for urgent caution in the use of colistin in animal husbandry.

A gold nanoparticles-assisted multiplex PCR assay for simultaneous detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

2020 ◽  
Vol 12 (2) ◽  
pp. 212-217 ◽  
Author(s):  
Juan Du ◽  
Shujing Wu ◽  
Liyuan Niu ◽  
Junguang Li ◽  
Dianbo Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gold Nanoparticles ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Pcr Product

Unfunctionalized flower-shaped AuNPs is used as colorimetric sensor for PCR product detection by naked eyes.

scholarly journals Rapid Identification of Different Escherichia coli Sequence Type 131 Clades

2017 ◽  
Vol 61 (8) ◽  
Author(s):  
Yasufumi Matsumura ◽  
Johann D. D. Pitout ◽  
Gisele Peirano ◽  
Rebekah DeVinney ◽  
Taro Noguchi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Epidemiological Studies ◽  
Rapid Identification ◽  
Sequence Type ◽  
Fluoroquinolone Resistance ◽  
Pcr Assay ◽  
Content Type ◽  
E Coli ◽  
Multiplex Pcr Assay ◽  
Clade C

ABSTRACT Escherichia coli sequence type 131 (ST131) is a pandemic clonal lineage that is responsible for the global increase in fluoroquinolone resistance and extended-spectrum-β-lactamase (ESBL) producers. The members of ST131 clade C, especially subclades C2 and C1-M27, are associated with ESBLs. We developed a multiplex conventional PCR assay with the ability to detect all ST131 clades (A, B, and C), as well as C subclades (C1-M27, C1-nM27 [C1-non-M27], and C2). To validate the assay, we used 80 ST131 global isolates that had been fully sequenced. We then used the assay to define the prevalence of each clade in two Japanese collections consisting of 460 ESBL-producing E. coli ST131 (2001-12) and 329 E. coli isolates from extraintestinal sites (ExPEC) (2014). The assay correctly identified the different clades in all 80 global isolates: clades A (n = 12), B (n = 12), and C, including subclades C1-M27 (n = 16), C1-nM27 (n = 20), C2 (n = 17), and other C (n = 3). The assay also detected all 565 ST131 isolates in both collections without any false positives. Isolates from clades A (n = 54), B (n = 23), and C (n = 483) corresponded to the O serotypes and the fimH types of O16-H41, O25b-H22, and O25b-H30, respectively. Of the 483 clade C isolates, C1-M27 was the most common subclade (36%), followed by C1-nM27 (32%) and C2 (15%). The C1-M27 subclade with bla CTX-M-27 became especially prominent after 2009. Our novel multiplex PCR assay revealed the predominance of the C1-M27 subclade in recent Japanese ESBL-producing E. coli isolates and is a promising tool for epidemiological studies of ST131.

Detection of fluoroquinolone resistance level in clinical canine and feline Escherichia coli pathogens using rapid real-time PCR assay

2009 ◽  
Vol 139 (3-4) ◽  
pp. 379-385 ◽  
Author(s):  
Bashar W. Shaheen ◽  
Chengming Wang ◽  
Calvin M. Johnson ◽  
Bernhard Kaltenboeck ◽  
Dawn M. Boothe
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Fluoroquinolone Resistance ◽  
Pcr Assay ◽  
Resistance Level

Polymerase Chain Reaction Assay for Detection of Escherichia coli O157:H7 and Escherichia coli O157:H−

2002 ◽  
Vol 65 (1) ◽  
pp. 5-11 ◽  
Author(s):  
TAKAHISA MIYAMOTO ◽  
NATSUKO ICHIOKA ◽  
CHIE SASAKI ◽  
HIROSHI KOBAYASHI ◽  
KEN-ICHI HONJOH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Pcr Assay ◽  
E Coli ◽  
Pcr Product ◽  
Genomic Dnas ◽  
Polymerase Chain

The DNA band patterns generated by polymerase chain reaction (PCR) using the du2 primer and template DNAs from various strains of Escherichia coli and non–E. coli bacteria were compared. Among three to five prominent bands produced, the three bands at about 1.8, 2.7, and 5.0 kb were detected in all of the E. coli O157 strains tested. Some nonpathogenic E. coli and all pathogenic E. coli except E. coli O157 showed bands at 1.8 and 5.0 kb. It seems that the band at 2.7 kb is specific to E. coli O157. Sequence analysis of the 2.7-kb PCR product revealed the presence of a DNA sequence specific to E. coli O157:H− and E. coli O157:H7. Since the DNA sequence from base 15 to base 1008 of the PCR product seems to be specific to E. coli O157, a PCR assay was carried out with various bacterial genomic DNAs and O157-FHC1 and O157-FHC2 primers that amplified the region between base 23 and base 994 of the 2.7-kb PCR product. A single band at 970 bp was clearly detected in all of the strains of E. coli O157:H− and E. coli O157:H7 tested. However, no band was amplified from template DNAs from other bacteria, including both nonpathogenic and pathogenic E. coli except E. coli O157. All raw meats inoculated with E. coli O157:H7 at 3 × 100 to 3.5 × 102 CFU/25 g were positive both for our PCR assay after cultivation in mEC-N broth at 42°C for 18 h and for the conventional cultural method.

scholarly journals Molecular Detection of Diarrheagenic Escherichia coli from Children with Acute Diarrhea in Tertiary Care Hospitals of Dhaka, Bangladesh.

2013 ◽  
Vol 5 (2) ◽  
pp. 59-66 ◽  
Author(s):  
Sushmita Roy ◽  
SM Shamsuzzaman ◽  
Kazi Z Mamun
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Acute Diarrhea ◽  
Tertiary Care ◽  
Medical Science ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Stool Samples

Objective: Multiplex PCR assay was used for diagnosis of diarrheagenic Escherichia coli (DEC) in stool samples of children (under 5 years) with acute diarrhea.  Methods: Samples were collected from January 2011 to December 2011, from Dhaka Medical College Hospital and Dhaka Shishu Hospital. Multiplex PCR with five specific primer pairs to detect enteropathogenic E. coli (eae, bfp), enterotoxigenic E. coli (lt, st) and enteroaggregative E. coli (aat) were used. However, enteroinvasive E. coli, enterohemorrhagicE. coli and diffusely adhererentE. coli were not sought. Result: In total, 135 (67.5%) E. coli were isolated from 200 stool samples. The prevalence of DEC was 68 (34%). Among DEC, most frequently isolated pathotype was EPEC 40 (58.82%), followed by ETEC 24 (35.29%) and EAggEC 18 (26.47%). Among the EPEC, 5 (12.5%) were typical EPEC. Among the 68 DEC positive cases, 22 samples contained more than one pathogenic gene in various combinations. Among the combination of DEC, EPEC+ETEC combination was 6 (27.27%) followed by ETEC+EAggEC 4 (18.18%), EPEC+EAggEC and ETEC+EPEC+EAggEC were both in 3 (13.6%). Conclusion:This study shows that DEC is a common cause of childhood diarrhea in Dhaka city of Bangladesh. By using multiplex PCR assay, DEC can be diagnosed in one PCR reaction that makes a conclusive diagnosis of diarrhea. DOI: http://dx.doi.org/10.3126/ajms.v5i2.8576 Asian Journal of Medical Science, Volume-5(2) 2014: 59-66

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