1,721,257 research outputs found
Mass spectrometry applications in microbiology beyond microbe identification: progress and potential
No full textIntroduction: Mass spectrometry (MS), particularly MALDI-time of flight (MALDI-TOF), has become a routine tool for microorganism identification in clinical microbiology laboratories in the last five years. The use of MALDI-TOF MS has accelerated laboratory analysis, thus providing accurate species-level information with very short turnaround times. Areas covered: Beyond microbe identification, MALDI-TOF MS offers great opportunities for fast strain typing and detection of antimicrobial susceptibility/resistance in both bacterial and fungal organisms. Drawing on evidence from PubMed literature searches, clinical microbiology laboratory experience, and the authors’ opinions, this review summarizes recent significant advances and ongoing challenges in these areas. Expert commentary: In the near future, it is expected that the implementation of new analytical algorithms, automation of procedures, and refinement of assays will enhance the clinical and epidemiological usefulness of MALDI-TOF and other MS technologies
MALDI-TOF mass spectrometry: any use for Aspergilli?
No full textRecently, relentless efforts to develop rapid, cost-effective, and reliable laboratory methods for daily diagnosis of fungal diseases such as aspergillosis appear to be materialized in the relatively new, but revolutionary matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS) technology. As for Aspergilli, MALDI-TOF MS profiling of isolates growing in culture--characteristic protein spectra are obtainable by means of simple and reproducible preanalytical and analytical procedures--ensures that single species within the different sections or complexes can be easily and accurately identified, including species that are morphologically and phylogenetically similar to each other. Thus, resort to longer and more onerous molecular biology techniques is restricted to those cases for which no spectra in the reference fungal database or library are available at the time of analysis. However, it is necessary to interrogate reference libraries composed of spectra that have been obtained using procedures similar to those used to obtain the test isolate's mass spectrum, as well as to continuously update these libraries for enriching them with fungal strains/species not (or not well) represented in their current versions. Compared to mold identification, very limited work was reported on the use of MALDI-TOF MS to perform strain typing or antifungal susceptibility testing for Aspergilli. If these complementing areas will be potentiated in the near future, MALDI-TOF MS could effectively support the clinical microbiology/mycology laboratory in its primary role of assisting either infection control specialists or physicians for the diagnosis and treatment of aspergillosis
Identification of Molds by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry
No full textAlthough to a lesser extent than diagnostic bacteriology, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently revolutionized the diagnostic mycology workflow. With regard to filamentous fungi (or molds), the precise recognition of pathogenic species is important for rapid diagnosis and appropriate treatment, especially for invasive diseases. This review summarizes the current experience with MALDI-TOF MS-based identification of common and uncommon mold species of Aspergillus, Fusarium, Mucorales, dimorphic fungi, and dermatophytes. This experience clearly shows that MALDI-TOF MS holds promise as a fast and accurate identification tool, particularly with common species or typical strains of filamentous fungi
The future of fungal susceptibility testing
No full textThe antifungal treatment failures and the emergence of resistant fungal strains have stimulated the need for reproducible and clinically relevant antifungal susceptibility testing (AFST). While the standard reference methods are not intended for routine use, commercial methods are widely used for performing AFST. However, to accelerate AFST and to improve the detection of antifungal resistance, which is the most challenging goal of AFST, novel assays have been developed. Following brief drug exposures of fungal cells, the new antifungal susceptibility end points seem to provide a reliable means of identifying fungal isolates, which harbor mutations that have been associated with antifungal resistance. This article summarizes the recent progress in AFST that is destined to enhance its clinical utility in the near future
Antifungal drug resistance among Candida species: mechanisms and clinical impact
No full textThe epidemiology of Candida infections has changed in recent years. Although Candida albicans is still the main cause of invasive candidiasis in most clinical settings, a substantial proportion of patients is now infected with non-albicans Candida species. The various Candida species vary in their susceptibility to the most commonly used antifungal agents, and the intrinsic resistance to antifungal therapy seen in some species, along with the development of acquired resistance during treatment in others, is becoming a major problem in the management of Candida infection. A better understanding of the mechanisms and clinical impact of antifungal drug resistance is essential for the efficient treatment of patients with Candida infection and for improving treatment outcomes. Herein, we report resistance to the azoles and echinocandins among Candida species
Looking for appropriateness in the cure of mixed vaginitis: the role of fenticonazole as an empiric treatment
No full textMixed vaginitis is defined as the simultaneous presence of at least two different vaginal pathogens, both contributing to an abnormal vaginal milieu leading to signs and symptoms. Pathogen coinfection occurs frequently in women with vaginitis, and both coinfection and mixed vaginitis have relevant clinical and therapeutic implications. Fenticonazole, an imidazole derivative with a broad spectrum of antimycotic and antimicrobial activity, appears at least as effective as other topical antifungals in the treatment of vulvovaginal candidiasis and can also have a major role in the treatment of mixed infections or coinfections of the lower genital tract. This paper will address the current role of topical fenticonazole as an empiric treatment of vulvovaginal infections, with a focus on the effectiveness in the treatment of mixed vaginitis and the possible implications of this
Development of a yeast retrotransposon-based system useful for screening of potentially active anti-retroviral compounds
No full textWe have developed a new system suitable for easy inexpensive screening of substances potentially active against HIV and other retroviruses. The system evaluates the ability of substances to inhibit the high-frequency induced retrotransposition of the yeast transposable element Ty917, and is based on an engineered version of the retrotransposon which carries a dominant selectable marker useful for scoring transposition events. The system performance was evaluated using two nucleoside analogues of proved anti-HIV activity, 3'-azido-3'deoxythymidine (AZT) and 2',3'-dideoxycytidine (ddC). Both substances were able to inhibit Ty917 transposition and could have been detected as potentially active antiretroviral drugs using the new screening system
Diffuse cutaneous candidiasis in a dog. Diagnosis by PCR-REA
No full textThe authors describe a clinical case of cutaneous candidiasis in a dog with dermatological lesions, characterized by persistent alopecia, crusts, ulcers and scales. Predisposing factors such as the use of corticosteroids, the concomitan presence of an autoimmune disease (pemphigus foliaceus) and an infection of ehrlichiosis caused by Ehrlichia canis were observed. Histopathological findings included signs of orthokeratotic hyperkeratosis, moderate follicular keratosis and light epidermic acanthosis. The reactive process included an infiltrative superficial dermatitis and a mural folliculitis with prevalent participation of macrophages and lymphocytes. The application of PCR-Restriction Enzyme Analysis (REA) method on cutaneous specimens in veterinary medicine is an extremely interesting diagnostic tool. Its use, together with other techniques, such as mycologic, cytologic and histological examinations, allowed us to identify Candida albicans as aetiological agent in this particular case
New approaches for antifungal susceptibility testing
Full text linkBACKGROUND:
Invasive fungal diseases, including those caused by (multi)drug-resistant Candida and Aspergillus species, still represent global public health concerns. Information about the antifungal susceptibility testing (AFST) of fungal isolates must be quickly produced to help clinicians in administrating appropriate antifungal therapies. Unfortunately, reference AFST methods, albeit accurate, are labour-intensive and take several hours before patients' results can be available to the treating clinicians.
AIMS & SOURCES:
This review is a blend of evidence obtained from PubMed literature searches, clinical laboratory experience, and the author's opinions that is aimed to summarize recent significant advances and ongoing challenges in the AFST area.
CONTENT:
Particular attention is given to the new approaches based on genetic or phenotypic recognition of antifungal resistance that are destined to enhance the clinical usefulness of AFST in the next future. Following short-time exposures of fungal cells to antifungal drugs, new antifungal susceptibility endpoints have been established, as well as novel diagnostic assay platforms have been proposed for the genotyping assessment of fungal isolates with resistance-associated mutations. Overall, new approaches provide a rapid, reliable means of identifying those fungal isolates with phenotypically detectable acquired resistance mechanisms, independently from the clinical susceptibility categorization of the isolates as obtained in a classical AFST way.
IMPLICATIONS:
Despite holding promise as a surrogate diagnostic method to better direct antifungal therapy, the AFST approaches described in this review need to be evaluated in multicentre laboratory studies to enable their standardization and refinement
Diagnostic of fungal infections related to biofilms
No full textFungal biofilm-related infections, most notably those caused by the Candida and Aspergillus genera, need to be diagnosed accurately and rapidly to avoid often unfavorable outcomes. Despite diagnosis of these infections is still based on the traditional histopathology and culture, the use of newer, rapid methods has enormously enhanced the diagnostic capability of a modern clinical mycology laboratory. Thus, while accurate specieslevel identification of fungal isolates can be achieved with turnaround times considerably shortened, nucleic acid-based or antigen-based detection methods can be considered useful adjuncts for the diagnosis of invasive forms of candidiasis and aspergillosis. Furthermore, simple, reproducible, and fast methods have been developed to quantify biofilm production by fungal isolates in vitro. In this end, isolates can be categorized as low, moderate, or high biofilm-forming, and this categorization may reflect their differential response to the conventional antifungal therapy. By means of drug susceptibility testing performed on fungal biofilm-growing isolates, it is now possible to evaluate not only the activity of conventional antifungal agents, but also of novel anti-biofilm agents. Despite this, future diagnostic methods need to target specific biofilm components/molecules, in order to provide a direct proof of the presence of this growth phenotype on the site of infection. In the meantime, our knowledge of the processes underlying the adaptive drug resistance within the biofilm has put into evidence biofilm-specific molecules that could be potentially helpful as therapeutic targets. Surely, the successful management of clinically relevant fungal biofilms will rely upon the advancement and/or refinement of these approaches
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