scholarly journals SCA Medium: A New Culture Medium for the Isolation of All Candida auris Clades

2021 ◽  
Vol 7 (6) ◽  
pp. 433
Author(s):  
Ahmad Ibrahim ◽  
Lucie Peyclit ◽  
Rim Abdallah ◽  
Saber Khelaifia ◽  
Amanda Chamieh ◽  
...  
Keyword(s):  
Culture Medium ◽  
Limit Of Detection ◽  
Bacterial Species ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Candida Auris ◽  
Phenotypic Profile ◽  
Stool Samples ◽  
Selective Culture Medium

Candida auris is an emerging multidrug-resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation are necessary for diagnosis and containing its spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection ranging between 101 and 102 CFU/mL. The 100% specificity of SCA (specific C. auris) medium is confirmed on a set of 135 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, allowing the latter to study its phenotypic profile.

scholarly journals SCA Medium: A New Culture Medium for the Isolation of all Candida auris Clade

2021 ◽  
Author(s):  
Ahmad Ibrahim ◽  
Lucie Peyclit ◽  
Rim Abdallah ◽  
Saber Khelaifia ◽  
Amanda Chamieh ◽  
...  
Keyword(s):  
Culture Medium ◽  
Limit Of Detection ◽  
Bacterial Species ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Candida Auris ◽  
Phenotypic Profile ◽  
Stool Samples ◽  
Selective Culture Medium

Candida auris is an emerging multidrug resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation is necessary for its diagnosis and containing spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection of 102 CFU/ml. The 100% specificity of SCA (Specific C. auris) medium is confirmed on a set of 134 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, and allows studying its phenotypic profile.

scholarly journals Rapid identification of health care–associated infections with an integrated fluorescence anisotropy system

2016 ◽  
Vol 2 (5) ◽  
pp. e1600300 ◽  
Author(s):  
Ki Soo Park ◽  
Chen-Han Huang ◽  
Kyungheon Lee ◽  
Yeong-Eun Yoo ◽  
Cesar M. Castro ◽  
...  
Keyword(s):  
Health Care ◽  
Fluorescence Anisotropy ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Integrated System ◽  
Clinical Samples ◽  
Drug Resistant ◽  
Polarization Anisotropy ◽  
Major Health ◽  
Health Care Associated Infections

Health care–associated infections (HAIs) and drug-resistant pathogens have become a major health care issue with millions of reported cases every year. Advanced diagnostics would allow clinicians to more quickly determine the most effective treatment, reduce the nonspecific use of broad-spectrum antimicrobials, and facilitate enrollment in new antibiotic treatments. We present a new integrated system, polarization anisotropy diagnostics (PAD), for rapid detection of HAI pathogens. The PAD uses changes of fluorescence anisotropy when detection probes recognize target bacterial nucleic acids. The technology is inherently robust against environmental noise and economically scalable for parallel measurements. The assay is fast (2 hours) and performed on-site in a single-tube format. When applied to clinical samples obtained from interventional procedures, the PAD determined the overall bacterial burden, differentiated HAI bacterial species, and identified drug resistance and virulence status. The PAD system holds promise as a powerful tool for near-patient, rapid HAI testing.

scholarly journals Detection of Candida albicans DNA from blood samples using a novel electrochemical assay

2011 ◽  
Vol 60 (4) ◽  
pp. 467-471 ◽  
Author(s):  
Alastair Muir ◽  
Gordon Forrest ◽  
John Clarkson ◽  
Alan Wheals
Keyword(s):  
Genomic Dna ◽  
Limit Of Detection ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Electrochemical Assay ◽  
Blood Samples ◽  
Resistant Species ◽  
Life Threatening ◽  
Appropriate Therapy ◽  
Species Specific

The genus Candida contains a number of yeast species which are opportunistic pathogens and are associated with life-threatening infections in immunocompromised individuals. Provision of appropriate therapy relies on the rapid identification of the infecting species, and existing methods of identifying Candida species in clinical samples are time and resource intensive and are not always specific enough to differentiate between drug-susceptible and drug-resistant species. We have previously developed a system for the rapid detection of yeast pathogens in clinical samples using PCR followed by hybridization with a suite of five species-specific, electrochemically labelled DNA probes. The limit of detection of the assay was shown to be 37 fg (∼1 genome) per reaction using extracted genomic DNA. We carried out a study to test the limit of detection of one of the probes, CA PR3, using blood samples from a healthy donor that were spiked with genomic DNA or with C. albicans cells. Our results demonstrated a limit of detection of 37 fg (ml blood)−1 (∼1 genome ml−1) using extracted DNA or 10 c.f.u. (ml blood)−1 using C. albicans cells, indicating that the assay is capable of detecting C. albicans nucleic acid at levels that are encountered in clinical samples.

scholarly journals Procalcitonin Detection in Veterinary Species: Investigation of Commercial ELISA Kits

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1511
Author(s):  
Federica Battaglia ◽  
Valentina Meucci ◽  
Rosalba Tognetti ◽  
Francesca Bonelli ◽  
Micaela Sgorbini ◽  
...  
Keyword(s):  
Veterinary Medicine ◽  
Recombinant Proteins ◽  
Limit Of Detection ◽  
Rapid Identification ◽  
Human Medicine ◽  
Clinical Samples ◽  
Reference Ranges ◽  
Plasma Samples ◽  
Elisa Kit ◽  
Species Specific

In human medicine, procalcitonin (PCT), the precursor of calcitonin, is used for the rapid identification of the origin and severity of sepsis. In veterinary medicine, PCT has been studied in horses, cattle, and dogs, but the use of PCT in diagnostic and/or prognostic settings is not possible because of the lack of validated assays to obtain reference ranges. The aim of the present study was the investigation of commercially available ELISA kits for the detection of canine and equine PCT in plasma samples. Validation of the ELISA kits was performed by using species-specific recombinant proteins spiked both in plasma and buffer samples; linearity, limit of detection (LOD), recovery, and intra-assay and inter-assay variability were calculated. Moreover, clinical samples obtained from sick and healthy animals were also analyzed with the tested kits. Canine PCT was measured with a recombinant canine and a canine PCT ELISA kit. Equine PCT was measured with an equine and a human ELISA PCT kit. Our data demonstrate that the canine recombinant PCT ELISA kit can be used to measure canine PCT in plasma samples, showing an intra-assay and inter-assay coefficient of variation less than 20% and a LOD of 11 pg/mL, whereas the present results do not support the use of the canine PCT ELISA kit. The human PCT ELISA kit is suitable to detect equine PCT with a LOD of 56 ng/mL, whereas the equine PCT ELISA kit did not detect recombinant equine PCT.

scholarly journals A Selective Culture Medium for Screening Ceftazidime-Avibactam Resistance in Enterobacterales and Pseudomonas aeruginosa

2020 ◽  
Vol 58 (9) ◽  
Author(s):  
Mustafa Sadek ◽  
Laurent Poirel ◽  
Camille Tinguely ◽  
Patrice Nordmann
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Lower Limit ◽  
Culture Medium ◽  
Limit Of Detection ◽  
Resistance Mechanisms ◽  
Gram Negative Bacteria ◽  
Excellent Performance ◽  
Gram Negative ◽  
Content Type ◽  
Selective Culture Medium

ABSTRACT The SuperCAZ/AVI medium was developed for screening ceftazidime-avibactam (CZA) resistance among Gram-negative bacteria (Enterobacterales and Pseudomonas aeruginosa). It was evaluated using 50 CZA-susceptible and 42 CZA-resistant Gram-negative isolates. Its sensitivity and specificity of detection were 100%. Excellent performance of the medium was also observed by testing spiked stools, with the lower limit of detection ranging from 101 to 102 CFU/ml. This screening medium provides the opportunity to detect CZA-resistant isolates regardless of their resistance mechanisms.

scholarly journals Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

2020 ◽  
Author(s):  
Jinghua Ruan ◽  
Wujun Wang ◽  
Tiying Zhang ◽  
Teng Zheng ◽  
Jing Zheng ◽  
...  
Keyword(s):  
Real Time ◽  
Limit Of Detection ◽  
Bacterial Species ◽  
High Specificity ◽  
Clinical Samples ◽  
Amplification Efficiency ◽  
Salmonella Spp ◽  
Specificity And Sensitivity ◽  
Qpcr Method ◽  
Real Time Qpcr

Abstract Salmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratiafonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/SL and 145 copies/SL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.

scholarly journals Molecular epidemiology of carbapenem resistant Enterobacteriaceae in Sri Lanka: First report of KPC-producing Klebsiella pneumoniae

2019 ◽  
Author(s):  
Wirittamulla Gamage Maheshika Kumudunie ◽  
Wijesooriya Rathnayaka Pathirennehalage Lakmini Inoka Wijesooriya ◽  
Kalubowilage Dhananja Namalie ◽  
Narapity Pathirannehalage Sunil-Chandra ◽  
Yasanandana Supunsiri Wijayasinghe
Keyword(s):  
Sri Lanka ◽  
Tertiary Care ◽  
Carbapenem Resistance ◽  
Epidemiological Data ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Care Hospital ◽  
Major Mechanism

Abstract Background Extended spectrum β-lactamase producing Enterobacteriaceae (ESBL-PE) and carbapenamase producing Enterocacteriaceae (CPE) are widely disseminated globally creating a huge public health threat. Even though incidence of multidrug-resistant Enterobacteriaceae is rapidly growing, the epidemiological data regarding the occurrence of CRE in Sri Lanka is scarce. In this study, we determined the prevalence of ESBP-PE and CRE and the genetic determinants of CPE. Methods A total of 593 clinically significant Enterobacteriaceae was isolated from different clinical samples (urine, pus/wound, respiratory, blood, and other sterile specimens) at a tertiary care hospital in Sri Lanka from December 2017 – February 2018. Antimicrobial susceptibility and identification of ESBL-PE, CRE were done by disc diffusion method. CRE were identified to species level using a rapid identification kit and carbapenemase production was determined by modified carbapenem inactivation method. The presence of blaKPC, blaNDM, blaOXA-48-like genes were detected by PCR. Results The overall prevalence of ESBL-PE and CRE were found to be 26.0% and 9.6%, respectively. The rate of ESBL-PE in different sample types ranged from 18.2% to 30.8% with the highest prevalence among uropathogenic Enterobacteriaceae. The occurrence of CRE ranged from 6.7% to 20.8% with the highest prevalence among respiratory Enterobacteriaceae. CRE species identified were K. pneumoniae (80.7%), E. coli (5.3%), C. freundii (7.0%), P. rettgeri (3.5%), E. cloacae (1.7%), and E. aerogenes (1.7%). The carbapenemase production was detected in 94.7% of CRE isolates. The carbapenemases found were OXA-48-like (88.9%), NDM (14.8%), and KPC (3.7%). Conclusions The prevalence of CRE in Sri Lanka is alarming. Carbapenamse production was the major mechanism of carbapenem resistance and K. pneumoniae was the predominant CRE. Presence of KPC enzyme was detected in addition to the previously reported NDM and OXA-48-like carbapenamases in Sri Lanka. The rapid spread of CPE, necessitates the prompt implementation of preventive measures in Sri Lanka.

scholarly journals Rapid identification of Nocardia cyriacigeorgica from a brain abscess patient using MALDI-TOF-MS

2020 ◽  
Vol 2020 (10) ◽  
Author(s):  
Mohammed AlMogbel ◽  
Mohammed AlBolbol ◽  
Noura Elkhizzi ◽  
Hisham AlAjlan ◽  
John Philip Hays ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Nocardia Cyriacigeorgica ◽  
Diagnostic Microbiology ◽  
Tof Ms

Abstract Nocardia cyriacigeorgica (N. cyriacigeorgica) is most frequently associated with human infections, including chronic bronchitis, pulmonary disease and brain abscesses. In general, N. cyriacigeorgica causes infections in immunocompromised individuals and has been reported in clinical samples worldwide. However, the isolation and speciation of N. cyriacigeorgica in the routine diagnostic microbiology laboratory are complicated and time consuming. Recent mass spectrometry techniques such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-TOF-MS) have been successfully integrated into many routine diagnostic microbiology laboratories, allowing for the rapid, accurate and simple identification and speciation of many different microorganisms, including difficult-to-identify bacterial species. Here, we present a case report of a 65-year-old female patient from the neurology ward of Prince Sultan Military Medical City in Riyadh, Saudi Arabia, who was infected with N. cyriacigeorgica. The bacterium was successfully identified by MALDI-TOF-MS, with species identification subsequently confirmed by sequence analysis of the 16S ribosomal RNA.

scholarly journals A TaqMan Probe-Based Real-Time PCR Assay for the Rapid Identification of the Emerging Multidrug-Resistant Pathogen Candida auris on the BD Max System

2019 ◽  
Vol 57 (7) ◽  
Author(s):  
Amorce Lima ◽  
Raymond Widen ◽  
Grant Vestal ◽  
Dominic Uy ◽  
Suzane Silbert
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Taqman Probe ◽  
Pcr Assay ◽  
Candida Auris ◽  
Closely Related Species ◽  
Content Type ◽  
Deep Wound

ABSTRACT Candida auris is an emerging multidrug-resistant fungal pathogen that has been associated with nosocomial bloodstream and deep wound infections causing a high mortality rate mainly in intensive care unit (ICU) patients. Laboratories currently rely on phenotypic testing using commercial automated systems for identification of yeasts; however, this technique has often led to misidentification of C. auris to other closely related species. We developed and validated a TaqMan-based real-time PCR assay on the BD Max platform targeting ribosomal DNA (rDNA) region nucleotide sequences to quickly and accurately test for C. auris infection from culture and clinical specimens. The assay is highly specific, reproducible, and sensitive, allowing detection of as low as 1 C. auris CFU per reaction within 3 h.

scholarly journals PCR Assay for Species-Specific Identification ofBacteroides thetaiotaomicron

2000 ◽  
Vol 38 (4) ◽  
pp. 1672-1675 ◽  
Author(s):  
Lee-Jene Teng ◽  
Po-Ren Hsueh ◽  
Jui-Chang Tsai ◽  
Feng-Lin Chiang ◽  
Ching-Yi Chen ◽  
...  
Keyword(s):  
Bacterial Species ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
Accurate Method ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Restriction Enzyme Digestion ◽  
Amplification Product ◽  
Dot Blot ◽  
Pcr Assay

Bacteroides thetaiotaomicron is the second most frequently encountered species of the anaerobes isolated from clinical specimens. We developed a PCR-based assay for the rapid identification of B. thetaiotaomicron. Specific primers were based on shared amplicons of about 1.2 kb generated from B. thetaiotaomicron by randomly amplified polymorphic DNA. This 1.2-kb fragment was sequenced and then used to design a set of PCR amplification primers. This PCR generated an amplification product of 721 bp, which was unique to all 65 isolates of B. thetaiotaomicron tested. There was no amplification with isolates of other bacterial species. Restriction enzyme digestion of the amplification product and dot blot hybridization further verified the specificity of the assay. These results suggest that this PCR assay targets a nucleotide sequence that is strongly conserved in B. thetaiotaomicron. This simple and rapid PCR assay provides a rapid and accurate method for identification of B. thetaiotaomicron and shows promise for the detection of B. thetaiotaomicron in clinical samples.

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