Evaluation of fluorescence in situ hybridisation (FISH) for the detection of fungi directly from blood cultures and cerebrospinal fluid from patients with suspected invasive mycoses

Silva, Roberto Moreira da; Neto, João Ricardo da Silva; Santos, Carla Silvana da Silva; Frickmann, Hagen; Poppert, Sven; Cruz, Kátia Santana; Koshikene, Daniela; Souza, João Vicente Braga de

doi:10.1186/s12941-015-0065-5

Cited by 18 publications

(18 citation statements)

References 31 publications

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“…For the rapid diagnosis of Candida bloodstream infections, technologies such as MALDI-TOF allow the direct identification of bacteria from positive blood cultures whereas yeasts are poorly identified using this technique [76,77]. New technologies (i.e., fluorescence in situ hybridization) [78] capable of identifying Candida spp. in blood cultures within a few hours are highly attractive, but clinical studies are needed to establish the clinical benefits of these diagnostic tools and how to introduce them into the diagnostic work-up.…”

Section: What Are Remaining Areas Of Uncertainty? Invasive Candidiasismentioning

confidence: 99%

Intensive care medicine research agenda on invasive fungal infection in critically ill patients

et al. 2017

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Purpose:To describe concisely the current standards of care, major recent advances, common beliefs that have been contradicted by recent trials, areas of uncertainty, and clinical studies that need to be performed over the next decade and their expected outcomes with regard to Candida and Aspergillus infections in non-neutropenic patients in the ICU setting. Methods:A systematic review of the medical literature taking account of national and international guidelines and expert opinion. Results:Severe invasive fungal infections (IFIs) are becoming increasingly frequent in critically ill patients. Approximately 80% of IFIs are due to Candida spp. and 0.3-19% to Aspergillus spp. Recent observations emphasize the necessity of building a worldwide sentinel network to monitor the emergence of new fungal species and changes in susceptibility. Robust data on the attributable mortality are essential for the design of clinical studies with mortality endpoints. Although early antifungal therapy for Candida has been recommended in patients with risk factors, sepsis of unknown cause, and positive Candida serum biomarkers [β-1 → 3-d-glucan (BDG) and Candida albicans germ tube antibody (CAGTA)], its usefulness and influence on outcome need to be confirmed. Future studies may specifically address the optimal diagnostic and therapeutic strategies for patients with abdominal candidiasis. Better knowledge of the pharmacokinetics of antifungal molecules and tissue penetration is a key issue for intensivists. Regarding invasive aspergillosis, further investigation is needed to determine its incidence in the ICU, its relationship with influenza outbreaks, the clinical impact of rapid diagnosis, and the significance of combination treatment. Conclusions:Fundamental questions regarding IFI have to be addressed over the next decade. The clinical studies described in this research agenda should provide a template and set priorities for the clinical investigations that need to be performed.

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Section: What Are Remaining Areas Of Uncertainty? Invasive Candidiasismentioning

confidence: 99%

Intensive care medicine research agenda on invasive fungal infection in critically ill patients

et al. 2017

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“…For example, spectrophotometric approaches have been used to detect Paracoccidioides brasiliensis [ 138 ], and more recently a combination of artificial intelligence and metabolomics was reported for paracoccidioidomycosis [ 139 ]. Fluorescence in situ hybridization methods have been described for Aspergillus and Candida species [ 140 , 141 ]. Matrix-assisted laser desorptionionization time of flight (MALDI-TOF) analysis now provides highly accurate and rapid identification strategies for Candida and other yeast species which are rapidly being adapted for filamentous fungi [ 109 , 142 ].…”

Section: Emerging Diagnostic Methodsmentioning

confidence: 99%

Biosensors and Diagnostics for Fungal Detection

Hussain

Malavia

Johnson

et al. 2020

JoF

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Early detection is critical to the successful treatment of life-threatening infections caused by fungal pathogens, as late diagnosis of systemic infection almost always equates with a poor prognosis. The field of fungal diagnostics has some tests that are relatively simple, rapid to perform and are potentially suitable at the point of care. However, there are also more complex high-technology methodologies that offer new opportunities regarding the scale and precision of fungal diagnosis, but may be more limited in their portability and affordability. Future developments in this field are increasingly incorporating new technologies provided by the use of new format biosensors. This overview provides a critical review of current fungal diagnostics and the development of new biophysical technologies that are being applied for selective new sensitive fungal biosensors to augment traditional diagnostic methodologies.

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“…FISH yöntemi ile, kültür ve fenotipik özelliklerin belirlenmesine gerek kalmadan etken mikroorganizmanın tanısı kısa sürede yapılabilmektedir. Duyarlılık ve özgüllüğü yüksek olan bu yöntemin, pratik ve kolay olmasının yanı sıra yaklaşık 90 dakika gibi kısa bir sürede tür düzeyinde tanımlamaya olanak sağlaması önemli avantajlardır 8,9 . Bu yöntemin, kan kültürlerinden Candida türlerinin doğru ve hızlı tanımlanmasında ve kandidemi ile ilgili mortalite ve morbiditeyi azaltacak etkin antifungal tedavinin uygulanmasında önemli olduğu rapor edilmiştir [10][11][12] .…”

Section: Gereç Ve Yöntemunclassified

“…Son yıllarda özellikle hızlı tanı için moleküler yöntemler geliştirilmiştir 6,7 . Yapılan çalışmalarda, pozitif kan kültürlerinde C.albicans, C.parapsilosis, C.tropicalis, C.glabrata ve C.krusei'nin tanımlanması için tasarlanmış, FDA onayı almış peptid nükleik asit fl oresan in situ hibridizasyon (PNA FISH) (Yeast Traffi c Light ® PNA FISH, AdvanDx) yönteminin kullanılabileceği bildirilmektedir 7,[8][9][10][11][12] . Bu çalışmada, kan kültürlerinde üreyen Candida türlerinin tanımlanmasında, PNA FISH yöntemin değerlendirilmesi amaçlanmıştır.…”

Section: Introductionunclassified

Evaluation of Peptide Nucleic Acid Fluorescent In Situ Hybridization (PNA FISH) Method in the Identification of Candida Species Isolated From Blood Cultures

Aydemir¹,

Koç

Atalay³

2016

Mikrobiyol Bul

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In recent years, increased number of patients who are hospitalized in intensive care units, received immunosuppressive therapy and treated with broad-spectrum antibiotics that can lead an increase in the incidence of systemic candidiasis. In these patients, the most common clinical manifestation is candidemia. Since the identification of Candida species isolated from blood cultures is time consuming by conventional (morphological and biochemical) methods, rapid, reliable and accurate methods are needed. For this purpose novel systems have been developed to identify the agent directly. The aim of this study was to evaluate the peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method for the identification of Candida species by comparing with the conventional methods. A total of 50 patients who were admitted to Erciyes University Medical Faculty Hospital clinics and followed with prediagnosis of systemic fungal infections whose blood cultures were positive for the yeasts between July 2011 and July 2012 were included in the study. The conventional identification of Candida isolates was performed by considering macroscopic and microscopic morphology, germ tube test, cycloheximide sensitivity, urease activity and carbohydrate assimilation patterns with API 20C AUX (bioMerieux, France) test. PNA FISH method was conducted by the use of a commercial kit namely Yeast Traffic Light(®) PNA FISH (AdvanDx, USA). According to morphological and biochemical characteristics (conventional methods), 19 (38%) out of 50 Candida isolates were identified as C.albicans, 12 (24%) as C.glabrata, five (10%) as C.parapsilosis, five (10%) as C.kefyr, four (8%) as C.krusei, two (4%) as C.guilliermondii, two (4%) as C.tropicalis and one (2%) as C.lusitaniae. On the other hand, 24 (48%) of the isolates were identified as C.albicans/C.parapsilosis (with green fluorescence), 16 (32%) as C.glabrata/C.krusei (with red fluorescence) and one (%2) as C.tropicalis (with yellow fluorescence) properly, however one C.tropicalis strain was misidentified as C.albicans by PNA FISH method. Other eight (16%) strains which were not presented in the evaluation panel of PNA FISH kit (5 C.kefyr, 2 C.guillermondii and 1 C.lusitaniae), gave no fluorescence and determined as other Candida spp. According to this, when the species that could be detected with the kit (C.albicans, C.parapsilosis, C.glabrata, C.krusei and C.tropicalis) were considered, the concordance rate with the conventional methods was determined as 97.6% (41/42) and the total evaluation rate for all the species was 84% (41/50). In conclusion, the most frequent isolated species from blood cultures in our hospital was C.albicans, followed by C.glabrata and C.parapsilosis. Since PNA FISH testing is a practical, reliable and rapid (resulted in 90 minutes) method for the identification of Candida strains at species level isolated from blood cultures, it was thought to be useful in routine laboratories. However, further comparative studies are required with large number of strains with the ...

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Evaluation of fluorescence in situ hybridisation (FISH) for the detection of fungi directly from blood cultures and cerebrospinal fluid from patients with suspected invasive mycoses

Cited by 18 publications

References 31 publications

Intensive care medicine research agenda on invasive fungal infection in critically ill patients

Intensive care medicine research agenda on invasive fungal infection in critically ill patients

Biosensors and Diagnostics for Fungal Detection

Evaluation of Peptide Nucleic Acid Fluorescent In Situ Hybridization (PNA FISH) Method in the Identification of Candida Species Isolated From Blood Cultures

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