Outbreak of Corynebacterium pseudodiphtheriticum Infection in Cystic Fibrosis Patients, France

Respiratory tract colonization with these bacteria may be common in this population

International Society of Human and Animal Mycology (ISHAM)-ITS reference DNA barcoding database - the quality controlled standard tool for routine identification of human and animal pathogenic fungi

Human and animal fungal pathogens are a growing threat worldwide leading to emerging infections and creating new risks for established ones. There is a growing need for a rapid and accurate identification of pathogens to enable early diagnosis and targeted antifungal therapy. Morphological and biochemical identification methods are time-consuming and require trained experts. Alternatively, molecular methods, such as DNA barcoding, a powerful and easy tool for rapid monophasic identification, offer a practical approach for species identification and less demanding in terms of taxonomical expertise. However, its wide-spread use is still limited by a lack of quality-controlled reference databases and the evolving recognition and definition of new fungal species/complexes. An international consortium of medical mycology laboratories was formed aiming to establish a quality controlled ITS database under the umbrella of the ISHAM working group on "DNA barcoding of human and animal pathogenic fungi." A new database, containing 2800 ITS sequences representing 421 fungal species, providing the medical community with a freely accessible tool at http://www.isham.org and http://its.mycologylab.org/ to rapidly and reliably identify most agents of mycoses, was established. The generated sequences included in the new database were used to evaluate the variation and overall utility of the ITS region for the identification of pathogenic fungi at intra-and interspecies level. The average intraspecies variation ranged from 0 to 2.25%. This highlighted selected pathogenic fungal species, such as the dermatophytes and emerging yeast, for which additional molecular methods/genetic markers are required for their reliable identification from clinical and veterinary specimens.This study was supported by an National Health and Medical Research Council of Australia (NH&MRC) grant [#APP1031952] to W Meyer, S Chen, V Robert, and D Ellis; CNPq [350338/2000-0] and FAPERJ [E-26/103.157/2011] grants to RM Zancope-Oliveira; CNPq [308011/2010-4] and FAPESP [2007/08575-1] Fundacao de Amparo Pesquisa do Estado de So Paulo (FAPESP) grants to AL Colombo; PEst-OE/BIA/UI4050/2014 from Fundacao para a Ciencia e Tecnologia (FCT) to C Pais; the Belgian Science Policy Office (Belspo) to BCCM/IHEM; the MEXBOL program of CONACyT-Mexico, [ref. number: 1228961 to ML Taylor and [122481] to C Toriello; the Institut Pasteur and Institut de Veil le Sanitaire to F Dromer and D Garcia-Hermoso; and the grants from the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) and the Fundacao de Amparo a Pesquisa do Estado de Goias (FAPEG) to CM de Almeida Soares and JA Parente Rocha. I Arthur would like to thank G Cherian, A Higgins and the staff of the Molecular Diagnostics Laboratory, Division of Microbiology and Infectious Diseases, Path West, QEII Medial Centre. Dromer would like to thank for the technical help of the sequencing facility and specifically that of I, Diancourt, A-S Delannoy-Vieillard, J-M Thiberge (Genotyping of Pathogens and Public Health, Institut Pasteur). RM Zancope-Oliveira would like to thank the Genomic/DNA Sequencing Platform at Fundacao Oswaldo Cruz-PDTIS/FIOCRUZ [RPT01A], Brazil for the sequencing. B Robbertse and CL Schoch acknowledge support from the Intramural Research Program of the NIH, National Library of Medicine. T Sorrell's work is funded by the NH&MRC of Australia; she is a Sydney Medical School Foundation Fellow.info:eu-repo/semantics/publishedVersio

Mould Routine Identification in the Clinical Laboratory by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry

BACKGROUND: MALDI-TOF MS recently emerged as a valuable identification tool for bacteria and yeasts and revolutionized the daily clinical laboratory routine. But it has not been established for routine mould identification. This study aimed to validate a standardized procedure for MALDI-TOF MS-based mould identification in clinical laboratory. MATERIALS AND METHODS: First, pre-extraction and extraction procedures were optimized. With this standardized procedure, a 143 mould strains reference spectra library was built. Then, the mould isolates cultured from sequential clinical samples were prospectively subjected to this MALDI-TOF MS based-identification assay. MALDI-TOF MS-based identification was considered correct if it was concordant with the phenotypic identification; otherwise, the gold standard was DNA sequence comparison-based identification. RESULTS: The optimized procedure comprised a culture on sabouraud-gentamicin-chloramphenicol agar followed by a chemical extraction of the fungal colonies with formic acid and acetonitril. The identification was done using a reference database built with references from at least four culture replicates. For five months, 197 clinical isolates were analyzed; 20 were excluded because they were not identified at the species level. MALDI-TOF MS-based approach correctly identified 87% (154/177) of the isolates analyzed in a routine clinical laboratory activity. It failed in 12% (21/177), whose species were not represented in the reference library. MALDI-TOF MS-based identification was correct in 154 out of the remaining 156 isolates. One Beauveria bassiana was not identified and one Rhizopus oryzae was misidentified as Mucor circinelloides. CONCLUSIONS: This work's seminal finding is that a standardized procedure can also be used for MALDI-TOF MS-based identification of a wide array of clinically relevant mould species. It thus makes it possible to identify moulds in the routine clinical laboratory setting and opens new avenues for the development of an integrated MALDI-TOF MS-based solution for the identification of any clinically relevant microorganism

MixInYeast: A Multicenter Study on Mixed Yeast Infections

Invasive candidiasis remains one of the most prevalent systemic mycoses, and several studies have documented the presence of mixed yeast (MY) infections. Here, we describe the epidemiology, clinical, and microbiological characteristics of MY infections causing invasive candidiasis in a multicenter prospective study. Thirty-four centers from 14 countries participated. Samples were collected in each center between April to September 2018, and they were sent to a reference center to confirm identification by sequencing methods and to perform antifungal susceptibility testing, according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST). A total of 6895 yeast cultures were identified and MY occurred in 150 cases (2.2%). Europe accounted for the highest number of centers, with an overall MY rate of 4.2% (118 out of 2840 yeast cultures). Of 122 MY cases, the most frequent combinations were Candida albicans/C. glabrata (42, 34.4%), C. albicans/C. parapsilosis (17, 14%), and C. glabrata/C. tropicalis (8, 6.5%). All Candida isolates were susceptible to amphotericin B, 6.4% were fluconazole-resistant, and two isolates (1.6%) were echinocandin-resistant. Accurate identification of the species involved in MY infections is essential to guide treatment decisions

Identification of medical fungi with MALDI-TOF MS : application of diagnosis to fungal diseases

L’avènement de la spectrométrie de masse de type MALDI-TOF a révolutionné la microbiologie en permettant l’identification précise des bactéries et des levures en seulement quelques minutes. En 2010, la MALDI-TOF SM n’était pas applicable à l’identification des champignons filamenteux. Un protocole d’identification générant des spectres interprétables de champignons filamenteux fut d’abord mis au point, suivi par le développement d’une architecture de banque originale. Enfin une banque de références solides, intégrant des références pour la majorité des espèces de moisissures impliquées en pathologie humaine, fut créée en collaboration avec le BCCM/IHEM. Les qualités d’identification de cette banque ont été démontrées non seulement à l’échelle localemais également à l’échelle nationale . Dans un second temps, nous sommes intéressés à l’identification des levures. Nous avons déterminé qu’une extraction complète est le meilleur protocole d’identification des levures en utilisant la banque de référence Bruker ™. Afin d’améliorer le diagnostic clinique,nous avons testé un protocole d’identification sur un séries de 6192 levures isolées de prélèvements cliniques reçus au laboratoire pendant un an. Enfin nous avons pu démontrer en utilisant nos « systèmes » d’identification par MALDI-TOF MS la sous-estimation de la diversité des espèces de moisissures et de levures isolées des prélèvements cliniques, lorsqu’on ne disposait que de techniques conventionnelles d’identification. La mise à disposition d’un tel outil ouvre de grandes perspectives dans le domaine de la mycologie, tant d’un point de vue épidémiologique que clinique.During the last decade, MALDI-TOF MS has revolutionized microbiology by enabling the accurate identification of bacteria and yeast in only few minutes1. At the beginning of this work in 2010, MALDI-TOF MS was not yet optimized for mold identification. To meet this need, we established an identification protocol to generate interpretable mold spectra. The structure of the reference database was developed taking intoaccount the heterogeneity of molds in culture. Finally, a comprehensive reference database,including references for the majority of molds encountered in human pathology, was created in collaboration with the BCCM/IHEM. Identification performance of this database was tested and validated at both a local scale and an international scale . We also determined the best-adapted pre-treatment protocol to identify yeasts in a routine setting. The protocol was tested on a panel of 6192 yeast isolates recovered from clinical samples submitted to our laboratory over the course of one year. Using our fungal identification system, we were able to identify morphologically similar species and highlight the underestimation of fungal pathogen diversity. The development of our MALDI-TOF MS-based fungal identification system presents numerous opportunities in the field of mycological, from both an epidemiological and clinical point of view. In subsequent studies, defining the clinical meaning of emerging species identified via MALDI-TOF MS will profoundly modify our perspective of fungal diseases

Etude de la faisabilité de l'identification des moisissures d'intérêt médical par spectrométrie de masse type MALDI TOF (Matrix-Assisted Laser Desorption/Ionisation-Time Of Flight)

AIX-MARSEILLE2-BU Pharmacie (130552105) / SudocSudocFranceF

Candida massiliensis sp. nov. Isolated from a Clinical Sample

International audienc

Identification of repositionable drugs with novel antimycotic activity by screening Prestwick Chemical Library against emerging invasive molds

International audienc