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

Lima, Amorce; Widen, Raymond; Vestal, Grant; Uy, Dominic; Silbert, Suzane

doi:10.1128/jcm.01604-18

Cited by 35 publications

(34 citation statements)

References 19 publications

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“…More recently, culture-independent methods have been developed for the detection of C. auris in few hours to allow the rapid identification of colonized patients. Both in-house [ 93 , 99 , 106 ] and commercial PCR-based assays [ 99 , 100 , 101 , 102 , 104 , 105 ] are available. These tests have performed well during clinical evaluations in which culture was used as the gold standard and have yielded >90% clinical sensitivities and specificities.…”

Section: Identification Of C Auris In Culture Isolmentioning

confidence: 99%

Candida auris: Epidemiology, Diagnosis, Pathogenesis, Antifungal Susceptibility, and Infection Control Measures to Combat the Spread of Infections in Healthcare Facilities

2021

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Candida auris, a recently recognized, often multidrug-resistant yeast, has become a significant fungal pathogen due to its ability to cause invasive infections and outbreaks in healthcare facilities which have been difficult to control and treat. The extraordinary abilities of C. auris to easily contaminate the environment around colonized patients and persist for long periods have recently resulted in major outbreaks in many countries. C. auris resists elimination by robust cleaning and other decontamination procedures, likely due to the formation of ‘dry’ biofilms. Susceptible hospitalized patients, particularly those with multiple comorbidities in intensive care settings, acquire C. auris rather easily from close contact with C. auris-infected patients, their environment, or the equipment used on colonized patients, often with fatal consequences. This review highlights the lessons learned from recent studies on the epidemiology, diagnosis, pathogenesis, susceptibility, and molecular basis of resistance to antifungal drugs and infection control measures to combat the spread of C. auris infections in healthcare facilities. Particular emphasis is given to interventions aiming to prevent new infections in healthcare facilities, including the screening of susceptible patients for colonization; the cleaning and decontamination of the environment, equipment, and colonized patients; and successful approaches to identify and treat infected patients, particularly during outbreaks.

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Section: Identification Of C Auris In Culture Isolmentioning

confidence: 99%

Candida auris: Epidemiology, Diagnosis, Pathogenesis, Antifungal Susceptibility, and Infection Control Measures to Combat the Spread of Infections in Healthcare Facilities

2021

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“…In the United States, the Taqman-based qPCR is currently the mostly widely used culture-independent test and is employed for C. auris surveillance at CDC and the Wadsworth Center in New York, as well as an increasing number of Antibiotic Resistance Laboratory Network (AR Lab Network) laboratories. Recent publications have described successes adapting the Taqman qPCR to the BD Max system (Becton Dickinson, NJ, USA), which automates the test and substantially reduces associated labor [19,24,25]. Additional promising culture-independent tests have been developed, although their performance with clinical samples is not yet known [26][27][28][29][30][31].…”

Section: Candida Auris Identificationmentioning

confidence: 99%

Candida auris: A Review of Recommendations for Detection and Control in Healthcare Settings

Cáceres

Forsberg

Welsh

et al. 2019

JoF

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Candida auris is an emerging multidrug-resistant fungal pathogen. Since first reported in 2009, C. auris has caused healthcare outbreaks around the world, often involving high mortality. Identification of C. auris has been a major challenge as many common conventional laboratory methods cannot accurately detect it. Early detection and implementation of infection control practices can prevent its spread. The aim of this review is to describe recommendations for the detection and control of C. auris in healthcare settings. most common misidentifications based on frequently used yeast identification methods [5,6]. However, efforts to improve C. auris identification methods have made substantial progress in the last few years. The development of a high-salt, high-temperature enrichment culture-based method has made it possible to reliably isolate C. auris from complex sample types [7][8][9]. Once an isolate is obtained, identification of C. auris can be efficiently accomplished with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. For MALDI-TOF identification, it is important to ensure C. auris is in the reference database [5,6,10]. The Bruker Biotyper (Bruker Daltonik GmbH, Bremen, Germany) and the VITEK MS (bioMérieux, Marcy, L'Etoile, France) include C. auris in their Research Use Only and certain versions of their FDA-approved system databases [3,5,11]. If MALDI-TOF is not available, laboratories can reliably identify an isolate by sequencing the D1-D2 region of the 28s rDNA or the internal transcribed spacer (ITS) regions of rDNA [12][13][14].

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“…The yeast suspension count was adjusted to 10 7 cells/mL. Ten-fold serial dilutions ranging from 10 6 to 10 0 CFU/mL were prepared to determine the analytical sensitivity of the assay [ 27 , 28 ]. To check the CFU count of viable C. auris in each dilution, 100 µL of each dilution was cultured on SDA plates.…”

Section: Methodsmentioning

confidence: 99%

“…When MALDI-TOF MS capability is unavailable, developing specific molecular identification methods based on DNA amplification is a potential aid to identifying C. auris . Recently, several in-house PCR- based methods, including real-time PCR assays with hydrolysis probes or melting curve analysis, have been introduced to detect this species [ 26 - 28 ]. The development of a specific quantitative PCR assay for C. auris can play an important role in the rapid and accurate identification of this species and is thus potentially useful in time-limited situations and diagnostic laboratories without access to advanced systems, such as MALDI-TOF MS.…”

Section: Introductionmentioning

confidence: 99%

Development a hydrolysis probe-based quantitative PCR assay for the specific detection and quantification of Candida auris

Jafarian¹,

Khodadadi²,

Badiee³

2020

CMM

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Background and Purpose: Candida auris is an emerging multidrug-resistant pathogen. The identification of this species with the conventional phenotypic or biochemical mycological methods may lead to misidentification. Molecular-based species-specific identification methods such as quantitative real-time polymerase chain reaction (qPCR) facilitate a more reliable identification of C. auris than mycological methods. Regarding this, the present study aimed to develop a hydrolysis probe-based qPCR assay for the rapid, accurate identification of C. auris. Materials and Methods: The internal transcribed spacer 2 regions in the nuclear ribosomal DNA of C. auris and other related yeasts were assayed to find a specific PCR target for C. auris. A 123-base-pair target was selected, and primers and a probe were designed for hydrolysis probe-based real-time PCR with TaqMan chemistry. Ten-fold serial dilutions of C. auris ranging from 106 to 100 CFU/mL were prepared to establish a standard curve to quantify the yeast. Results: The qPCR assay was able to identify and quantify C. auris with a detection limit of 1 C. auris CFU per reaction. Specificity was confirmed by the non-amplification of the sequences belonging to other Candida species, yeasts, molds, bacteria, or human DNAs. The standard curve of the assay showed a highly significant linearity between threshold values and dilution rates (R2=0.99; slope=−3.42). Conclusion: The applied qPCR assay facilitated the rapid and accurate identification and quantification of emerging opportunistic C. auris. Therefore, considering the promising test validation results, we succeeded to develop a rapid and accurate hydrolysis probe-based qPCR assay for the screening and identification of C. auris.

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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

Cited by 35 publications

References 19 publications

Candida auris: Epidemiology, Diagnosis, Pathogenesis, Antifungal Susceptibility, and Infection Control Measures to Combat the Spread of Infections in Healthcare Facilities

Candida auris: Epidemiology, Diagnosis, Pathogenesis, Antifungal Susceptibility, and Infection Control Measures to Combat the Spread of Infections in Healthcare Facilities

Candida auris: A Review of Recommendations for Detection and Control in Healthcare Settings

Development a hydrolysis probe-based quantitative PCR assay for the specific detection and quantification of Candida auris

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