scholarly journals Ultrasensitive and rapid identification of ESRI developer- and piperacillin/tazobactam-resistant Escherichia coli by the MALDIpiptaz test

Author(s):  
Ángel Rodríguez-Villodres ◽  
Lydia Gálvez Benítez ◽  
Manuel Arroyo ◽  
Gema Méndez ◽  
Luis Mancera ◽  
...  

Abstract The excessive use of piperacillin/tazobactam (P/T) has promoted the emergence of P/T-resistant Enterobacterales. We reported that in Escherichia coli, P/T contributes to the development of extended-spectrum resistance to β-lactam/β-lactamase inhibitor (BL/BLI) (ESRI) in isolates that are P/T susceptible but have low-level resistance to BL/BLI. Currently, the detection of P/T resistance relying on conventional methods is time-consuming. To overcome this issue, we developed a cost-effective test based on MALDI-MS technology, called MALDIpiptaz, which aims to detect P/T resistance and ESRI developers in E. coli. We used automated Clover MS Data Analysis software to analyse the protein profile spectra obtained by MALDI-MS from a collection of 248 E. coli isolates (91 P/T-resistant, 81 ESRI developers and 76 P/T-susceptible). This software allowed to preprocess all the spectra to build different peak matrices that were analysed by machine learning algorithms. We demonstrated that MALDIpiptaz can efficiently and rapidly (15 min) discriminate between P/T-resistant, ESRI developer and P/T-susceptible isolates and allowed the correct classification between ESRI developers from their isogenic resistance to P/T. The combination of excellent performance and cost-effectiveness are all desirable attributes, allowing the MALDIpiptaz test to be a useful tool for the rapid determination of P/T resistance in clinically relevant gram-negative bacteria.

2021 ◽  
Vol 9 ◽  
Author(s):  
Xiaofeng Lin ◽  
Yanqiu Mei ◽  
Chen He ◽  
Yan Luo ◽  
Min Yang ◽  
...  

Escherichia coli (E. coli) O157:H7 can cause many food safety incidents, which seriously affect human health and economic development. Therefore, the sensitive, accurate, and rapid determination of E. coli O157:H7 is of great significance for preventing the outbreak and spread of foodborne diseases. In this study, a carbon dots-Fe3O4 nanomaterial (CDs-Fe3O4)-based sensitive electrochemical biosensor for E. coli O157:H7 detection was developed. The CDs have good electrical conductivity, and the surface of carbon dots contains abundant carboxyl groups, which can be used to immobilize probe DNA. Meanwhile, the CDs can be used as a reducing agent to prepare CDs-Fe3O4 nanomaterial. The Fe3O4 nanomaterial can improve the performance of the electrochemical biosensor; it also can realize the recovery of CDs-Fe3O4 due to its magnetism. As expected, the electrochemical biosensor has excellent specificity of E. coli O157:H7 among other bacteria. The electrochemical biosensor also exhibited good performance for detecting E. coli O157:H7 with the detection range of 10–108 CFU/ml, and the detection limit of this electrochemical biosensor was 6.88 CFU/ml (3S/N). Furthermore, this electrochemical biosensor was successfully used for monitoring E. coli O157:H7 in milk and water samples, indicating that this electrochemical biosensor has good application prospect. More importantly, this research can provide a new idea for the detection of other bacteria and viruses.


1966 ◽  
Vol 12 (4) ◽  
pp. 699-702 ◽  
Author(s):  
D. E. Mahony ◽  
P. Chadwick

A means of determining the antibiotic sensitivity of a known pathogen in a mixture of organisms of varying antibiotic sensitivity is described. Such a technique involved the combination of a rapid antibiotic-sensitivity test in which the criterion of sensitivity was the inhibition of microcolony formation on the surface of agar plates containing antibiotic and the fluorescent-antibody reaction used in the rapid identification of bacteria. It was possible to determine the antibiotic sensitivity of enteropathogenic Escherichia coli in artificially mixed cultures within 5 hours by this method.


1979 ◽  
Vol 42 (2) ◽  
pp. 161-163 ◽  
Author(s):  
ROBERT M. TWEDT ◽  
BRENDA K. BOUTIN

Several coliform species other than Escherichia coli are often associated with and possibly responsible for acute and chronic diarrheal disease. Recent evidence suggests that non-Escherichia coli coliforms may be capable of colonizing the human intestine and producing enterotoxin(s) in high-yield. Whether these organisms are newly capable of causing disease because of infestation with extrachromosomal factors mediating pathogenicity or simply because of inherent pathogenic capabilities that have gone unrecognized, they pose a potential health hazard. Food, medical, and public health microbiologists should be aware that the non-E. coli coliforms contaminating foods may be potential enteropathogens. This possibility may make determination of their pathogenic capabilities even more important than identification of their taxonomic characteristics.


Author(s):  
Zorica Voşgan ◽  
Anca Domuţa ◽  
Stela Jelea ◽  
Lucia Mihălescu ◽  
Flavia Pop

Papers on beta- glucuronidase sensitivity and specificity for identifying Escherichia coli in sources of environment, food, water, etc. have been published since 1976. In this study we conducted a review of the incidence of E. coli β- glucuronidase -positive in goat milk, obtained by hand milking throughout the lactation: spring, summer, autumn. The presence of E. coli in milk is considered both as a health indicator and a pathogenic factor capable of causing food poisoning. The determination of the E. coli β-glucuronidase-positive was carried using TBX medium by cultivating colonies typical blue at 440C. The absence of E. coli in milk yielded during the spring, when the animal milking is done three times a day, was found in the performed analyses; the same was observed during fall, when the milk production is lower and the milking is done once a day. The load of E. coli β-glucuronidase-positive was averaging 66.67 CFU/ml of goat milk, during the middle lactation period (July-August), in conditions of higher temperature. During this period, milking is done in the mountain zone, where the transhumance of animals takes place in summer. The presence of the species E. coli was also confirmed by microscopic examination. Attention should be paid to hygiene and milk should be immediately cooled, during hot weather, as E. coli can be a source of food poisoning.  


2017 ◽  
Vol 78 (1) ◽  
pp. 155-159 ◽  
Author(s):  
M. Oliveira ◽  
D. Freire ◽  
N. M. Pedroso

Abstract The detection of pathogenic microorganisms in aquatic environments is extremely relevant in terms of public health. As these laboratorial methodologies are usually difficult, expensive and time-consuming, they are frequently replaced by the assessment of fecal indicator bacteria, such as Escherichia coli. This study aimed to assess the presence of E. coli in fecal samples from Neotropical otters, to evaluate its potential as fecal indicator to be applied to the determination of water microbiological quality in areas where otters’ populations are high. Twenty-six otter fecal samples, collected in Alto Paranapanema river basin, São Paulo State, Brazil, were analyzed for the presence of E. coli, using conventional bacteriological methods. Only 8 scat samples (30%) were E. coli positive, indicating that this microorganism is not a suitable fecal indicator to assess water fecal contamination by Neotropical otters, and should not be used to infer the presence of otter related pathogens in waters.


2017 ◽  
Vol 61 (8) ◽  
Author(s):  
Yasufumi Matsumura ◽  
Johann D. D. Pitout ◽  
Gisele Peirano ◽  
Rebekah DeVinney ◽  
Taro Noguchi ◽  
...  

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.


2001 ◽  
Vol 84 (2) ◽  
pp. 407-415
Author(s):  
Diane F Ekholm ◽  
Irvin N Hirshfield

Abstract Three methods to enumerate Escherichia coli in food were compared. They were based on AOAC methods using lauryl tryptose broth (LST) medium, LST-4-methylumbelliferyl-ß-D-glucuronide (MUG) medium, and a proposed method using regular LST in combination with E. coli (EC)–MUG medium. An efficacious and cost-effective method is needed that can detect E. Coli and does not produce false presumptive positives. We tested 170 cheeses, 40 frozen processed seafood samples, 210 tree nuts, and 40 other samples. The method of choice for enumerating E. Coli depends on the commodity itself. For a product, such as hard cheese or processed seafood, with a history of being negative for E. Coli and other lactose-fermenting organisms, the proposed method using regular LST/EC–MUG is a good choice. These samples were seldom presumptive positive in the primary LST medium. If gas was produced, EC–MUG was an effective secondary medium. No false positives (fluorescence) or negatives were detected in EC–MUG medium. For a product with a history of being positive for E. Coli and/or other lactose fermenting organisms, such as tree nutmeats or cheeses that are ripened by bacteria or mold, the method using LST–MUG is the method of choice. A presumptive positive in the LST–MUG medium was highly correlative with the biochemical tests that confirmed a sample contain E. Coli. For samples spiked with E. Coli, the results from each of these 3 methods were identical, and were consistent in enumerating E. Coli.


1996 ◽  
Vol 59 (6) ◽  
pp. 670-673 ◽  
Author(s):  
SHIU W. HUANG ◽  
TSUNG C. CHANG

A sandwich enzyme-immunoassay performed on plastic sticks was developed to specifically identify Escherichia coli O157. Colonies of test bacteria grown on tryptic soy agar were suspended in phosphate-buffered saline, heated in a 100°C water bath for 15 min, and incubated with the plastic sticks coated with a commercial preparation of anti-E. coli O157 antibodies at 37°C for 1.5 h. After incubation, the same antibodies labeled with peroxidase were used to produce the signal of antigen-antibody reaction. For 35 strains of E. coli O157 (among them 34 were E. coli O157:H7) tested, all produced strong reactions by the immunoassay. For 162 strains of E. coli with somatic antigens other than O157 and 38 strains of other genera tested, only one strain (Salmonella bietri) produced a false-positive reaction. The specificity and sensitivity of the immunostick assay were 100% (35/35) and 99.5% (199/200), respectively. The detection limit of the assay for E. coli O157:H7 (CCRC 15991) was about 105 CFU/ml. The method, which can be carried out within 3 h, is useful for rapid identification of suspect E. coli O157 isolated on selective media.


1978 ◽  
Vol 56 (9) ◽  
pp. 849-852 ◽  
Author(s):  
Ann D. E. Fraser ◽  
Hiroshi Yamazaki

We have developed a method for estimating the rates of synthesis and degradation of adenosine 3′,5′-cyclic monophosphate (cAMP) in Escherichia coli during balanced growth. Applying this method, we have found that an E. coli CRP− mutant 5333 (deficient for cAMP receptor protein) synthesizes cAMP about 25 times faster than does its CRP+ parent 1100. This accounts for the abnormally high intracellular and extracellular cAMP accumulation in 5333.


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