scholarly journals Detection of Ampicillin-Resistant E. coli Using Novel Nanoprobe-Combined Fluorescence In Situ Hybridization

Nanomaterials ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 750 ◽  
Author(s):  
Wang Sik Lee ◽  
Soohyun Lee ◽  
Taejoon Kang ◽  
Choong-Min Ryu ◽  
Jinyoung Jeong
Keyword(s):  
Antibiotic Resistance ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Antibiotic Resistance Gene ◽  
Sol Gel ◽  
Light Condition ◽  
Resistant Bacteria ◽  
Specific Gene ◽  
E Coli

Antibiotic-resistant bacteria present a global threat because the infections they cause are difficult to treat. Therefore, it is highly important to develop advanced methods for the identification of antibiotic resistance gene in the virulent bacteria. Here, we report the development of novel nanoprobes for fluorescence in situ hybridization (FISH) and the application of the nanoprobe to the detection of ampicillin-resistant Escherichia coli. The nanoprobe for FISH was synthesized by the modified sol–gel chemistry and the synthesized nanoprobe provided strong fluorescent signals and pH stability even under natural light condition. For the double-identification of bacteria species and ampicillin-resistance with a single probe in situ, the nanoprobes were conjugated to the two kinds of biotinylated probe DNAs; one for E. coli-species specific gene and the other for a drug-resistant gene. By using the nanoprobe-DNA conjugants, we successfully detected the ampicillin-resistant E. coli through the FISH technique. This result suggests the new insight into light stable FISH application of the nanoprobe for a pathogenic antibiotic-resistance bacterium.

scholarly journals Multiple NDM-5-Expressing Escherichia Coli Isolates From an Immunocompromised Pediatric Host

2020 ◽  
Vol 7 (2) ◽  
Author(s):  
Tim Flerlage ◽  
Jessica N Brazelton de Cardenas ◽  
Cherilyn D Garner ◽  
Nur A Hasan ◽  
Hiren Karathia ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antibiotic Resistance Gene ◽  
The United States ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Invasive Infection ◽  
Anticancer Chemotherapy ◽  
E Coli ◽  
Sequence Types

Abstract Background Genes conferring carbapenem resistance have disseminated worldwide among Gram-negative bacteria. Here we present longitudinal changes in clinically obtained Escherichia coli isolates from 1 immunocompromised pediatric patient. This report demonstrates potential for antibiotic resistance genes and plasmids to emerge over time in clinical isolates from patients receiving intensive anticancer chemotherapy and broad-spectrum antibiotics. Methods Thirty-three isolates obtained over 7 months from 1 patient were included. Clinical data were abstracted from the medical record. For each isolate, studies included phenotypic antibacterial resistance patterns, sequence typing, bacterial isolate sequencing, plasmid identification, and antibiotic resistance gene identification. Results Sites of isolation included blood, wound culture, and culture for surveillance purposes from the perianal area. Isolates were of 5 sequence types (STs). All were resistant to multiple classes of antibiotics; 23 (69.6%) were phenotypically resistant to all carbapenems. The blaNDM-5 gene was identified in 22 (67%) isolates, all of ST-167 and ST-940, and appeared to coincide with the presence of the IncFII and IncX3 plasmid. Conclusions We present unique microbiologic data from 33 multidrug-resistant E. coli isolates obtained over the course of 7 months from an individual patient in the United States. Two E. coli sequence types causing invasive infection in the same patient and harboring the blaNDM-5 gene, encoded on the IncX3 plasmid and the IncFII plasmid, were identified. This study highlights the emergence of multidrug-resistant bacteria on antibiotic therapy and the necessity of adequate neutrophil number and function in the clearance of bacteremia.

scholarly journals Analysis of Wastewater Reveals the Spread of Diverse Extended-Spectrum β-Lactamase-Producing E. coli Strains in uMgungundlovu District, South Africa

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 860
Author(s):  
Siyabonga N. Gumede ◽  
Akebe L. K. Abia ◽  
Daniel G. Amoako ◽  
Sabiha Y. Essack
Keyword(s):  
South Africa ◽  
Antibiotic Resistance ◽  
Membrane Filtration ◽  
Antibiotic Resistance Gene ◽  
Fourth Generation ◽  
Resistant Bacteria ◽  
Filtration Method ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum

Wastewater treatment plants (WWTPs) are major reservoirs of antibiotic-resistant bacteria (ARB), favouring antibiotic resistance genes (ARGs) interchange among bacteria and they can provide valuable information on ARB circulating in a community. This study characterised extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli from the influent and effluent of four WWTPs in uMgungundlovu District, KwaZulu-Natal, South Africa. E. coli was enumerated using the membrane filtration method and confirmed using the API 20E test and real-time polymerase chain reaction. ESBL-producers were phenotypically identified by their susceptibility to the third-generation cephalosporins using the disc diffusion and the double-disc synergy methods against cefotaxime (30 µg) with and without 10 µg clavulanic acid. Genotypic verification was by PCR of the TEM, SHV, and CTX-M genes. The clonality of isolates was assessed by ERIC-PCR. The highest E. coli count ranged between 1.1 × 105 (influent) and 4.3 × 103 CFU/mL (effluent). Eighty pure isolates were randomly selected, ten from the influent and effluent of each of the four WWTP. ESBLs were phenotypically confirmed in 49% (n = 39) of the isolates, of which 77% (n = 30) were genotypically confirmed. Seventy-three percent of the total isolates were multidrug-resistant (MDR). Only two isolates were susceptible to all antibiotics. Overall, resistance to first and second-generation cephalosporins was higher than to third and fourth generation cephalosporins. Also, 15% of the isolates were resistant to carbapenems. The CTX-M-type ESBL (67%; n = 20) was the most common ESBL antibiotic resistance gene (ARG) followed by TEM (57%; n = 17) and SHV-types (27%; n = 8). Also, a substantial number of isolates simultaneously carried all three ESBL genes. ERIC-PCR revealed a high diversity of isolates. The diversity of the isolates observed in the influent samples suggest the potential circulation of different ESBL-producing strains within the studied district, requiring a more comprehensive epidemiological study to prevent the spread of ESBL-producing bacteria within impoverished communities.

scholarly journals ASSESSMENT OF ANTIBIOTIC RESISTANCE AND BACTERIAL CONTAMINATION OF ICE SOLD IN CANTHO CITY, VIETNAM

2019 ◽  
Vol 57 (3B) ◽  
pp. 49
Author(s):  
Ngoc Thi Anh Tong
Keyword(s):  
Antibiotic Resistance ◽  
Bacterial Contamination ◽  
Antibiotic Resistance Gene ◽  
Resistant Bacteria ◽  
Antibiotics Resistance ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Can Tho City ◽  
The City

This study aimed to investigate the bacterial contamination of flake and cube ice being used dailyin the community. Thirty-one ice samples were collected from different areas in the city of Can Tho city, Vietnam. The enumeration of total aerobic mesophilic counts, the presence of coliforms and Escherichia coli (E. coli) and determination of antibiotics resistance of E. coli isolates were examined. The results indicated that total aerobic mesophilic counts ranged from 2.5 to 6.2 log CFU/mL and significant differences of total aerobic mesophilic counts were found between flake ice and cube ice (p < 0.05). Coliforms and E. coli were present on the ice samples of 93.55% and 58.06%, respectively. A total of 39 E. coli isolates were tested their resistance to 15 different antibiotics. The E. coli isolates of 74.36% were multi-resistance from three to thirteen antibiotics. The high prevalance was resistant to Ampicillin (79.49%), Cefotaxime (69.23%), Ceftazidime (46.15%), Tetracycline (56.41%), Sulfamethoxazole/Trimethoprime (46.15%), Colistin (20.51%), etc. As E. coli is an hygiene indicator and a candidate vehicle for the transfer of antibiotic resistance gene, it is highly recommended using clean and probable water in ice making as well as preventing the spread of antibiotic resistant bacteria.

scholarly journals Quantitative fluorescence in situ hybridization (FISH) of magnetically confined bacteria enables rapid determination of early-stage human bacteremia

2021 ◽  
Author(s):  
Min Seok Lee ◽  
Hwi Hyun ◽  
Inwon Park ◽  
Sungho Kim ◽  
Dong-Hyun Jang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Blood Culture ◽  
Fluorescence In Situ Hybridization ◽  
Rapid Determination ◽  
Early Stage ◽  
Culturable Bacteria ◽  
E Coli ◽  
Magnetically Confined

AbstractThe current diagnosis of bacteremia mainly uses blood culture, which is insufficient to offer rapid and quantitative determination of pathogens in blood. Here, we report a quantitative and sequential multiplexed fluorescence in situ hybridization in a microfluidic device (µFISH) that enables early and rapid (2-hour) diagnosis of bacteremia without prior blood culture. Mannose-binding lectin-coated magnetic nanoparticles enrich a broad range of pathogens, and µFISH enables identification and quantification of the magnetically confined bacteria. We detect Escherichia coli (E. coli) and measure their relative proportions to universal bacteria levels in the bacteremic blood of a porcine model and human whole blood collected from E. coli-infected patients, which was elusive with the conventional bacteremia diagnosis methods. Thus, µFISH can be used as a versatile tool to rapidly identify pathogens and further assess the number of both culturable and non-culturable bacteria in biological and environmental samples.

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