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White P. Lewis Alanio Alexandre Cruciani Mario Gorton Rebecca Millon Laurence Rickerts Volker Barnes Rosemary A. Donnelly J. Peter Loeffler Juergen 《Current fungal infection reports》2020,14(1):76-88
Current Fungal Infection Reports - This review has incorporated the knowledge and experience of the leads of each of the laboratory working parties of the fungal PCR initiative in order to provide... 相似文献
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Maud Gits-Muselli Marie-Noelle Peraldi Nathalie de Castro Véronique Delcey Jean Menotti Nicolas Guigue Samia Hamane Emmanuel Raffoux Anne Bergeron Sandrine Valade Jean-Michel Molina Stéphane Bretagne Alexandre Alanio 《PloS one》2015,10(5)
Pneumocystis pneumonia is a severe opportunistic infection in immunocompromised patients caused by the unusual fungus Pneumocystis jirovecii. Transmission is airborne, with both immunocompromised and immunocompetent individuals acting as a reservoir for the fungus. Numerous reports of outbreaks in renal transplant units demonstrate the need for valid genotyping methods to detect transmission of a given genotype. Here, we developed a short tandem repeat (STR)-based molecular typing method for P. jirovecii. We analyzed the P. jirovecii genome and selected six genomic STR markers located on different contigs of the genome. We then tested these markers in 106 P. jirovecii PCR-positive respiratory samples collected between October 2010 and November 2013 from 91 patients with various underlying medical conditions. Unique (one allele per marker) and multiple (more than one allele per marker) genotypes were observed in 34 (32%) and 72 (68%) samples, respectively. A genotype could be assigned to 55 samples (54 patients) and 61 different genotypes were identified in total with a discriminatory power of 0.992. Analysis of the allelic distribution of the six markers and minimum spanning tree analysis of the 61 genotypes identified a specific genotype (Gt21) in our hospital, which may have been transmitted between 10 patients including six renal transplant recipients. Our STR-based molecular typing method is a quick, cheap and reliable approach to genotype Pneumocystis jirovecii in hospital settings and is sensitive enough to detect minor genotypes, thus enabling the study of the transmission and pathophysiology of Pneumocystis pneumonia. 相似文献
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Gianina Ravenscroft Flora Nolent Sulekha Rajagopalan Ana M. Meireles Kevin J. Paavola Dominique Gaillard Elisabeth Alanio Michael Buckland Susan Arbuckle Michael Krivanek Jérome Maluenda Stephen Pannell Rebecca Gooding Royston W. Ong Richard J. Allcock Ellaine D.F. Carvalho Maria D.F. Carvalho Fernando Kok William S. Talbot Judith Melki Nigel G. Laing 《American journal of human genetics》2015,96(6):955-961
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Correlation Between Pneumocystis jirovecii Mitochondrial Genotypes and High and Low Fungal Loads Assessed by Single Nucleotide Primer Extension Assay and Quantitative Real‐Time PCR
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Alexandre Alanio Martine Olivi Odile Cabaret Françoise Foulet Anne‐Pauline Bellanger Laurence Millon Ana Berceanu Catherine Cordonnier Jean‐Marc Costa Stéphane Bretagne 《The Journal of eukaryotic microbiology》2015,62(5):650-656
We designed a single nucleotide primer extension (SNaPshot) assay for Pneumocystis jirovecii genotyping, targeting mt85 SNP of the mitochondrial large subunit ribosomal RNA locus, to improve minority allele detection. We then analyzed 133 consecutive bronchoalveolar lavage (BAL) fluids tested positive for P. jirovecii DNA by quantitative real‐time PCR, obtained from two hospitals in different locations (Hospital 1 [n = 95] and Hospital 2 [n = 38]). We detected three different alleles, either singly (mt85C: 39.1%; mt85T: 24.1%; mt85A: 9.8%) or together (27%), and an association between P. jirovecii mt85 genotype and the patient's place of hospitalization (p = 0.011). The lowest fungal loads (median = 0.82 × 103 copies/μl; range: 15–11 × 103) were associated with mt85A and the highest (median = 1.4 × 106 copies/μl; range: 17 × 103–1.3 × 107) with mt85CTA (p = 0.010). The ratios of the various alleles differed between the 36 mixed‐genotype samples. In tests of serial BALs (median: 20 d; range 4–525) from six patients with mixed genotypes, allele ratio changes were observed five times and genotype replacement once. Therefore, allele ratio changes seem more frequent than genotype replacement when using a SNaPshot assay more sensitive for detecting minority alleles than Sanger sequencing. Moreover, because microscopy detects only high fungal loads, the selection of microscopy‐positive samples may miss genotypes associated with low loads. 相似文献
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Alexandre Alanio Stéphane Romand Dominique Penso-Assathiany Françoise Foulet Françoise Botterel 《Mycopathologia》2011,171(1):61-65
We report a rare case of dermatophytosis due to Microsporum praecox in a 28-year-old female horse rider. The skin lesion was located on the right external malleolus. Microscopic examination
of skin scrapings revealed a dermatophyte which was also isolated in culture. The identification of M. praecox was confirmed by molecular biology (sequence analysis of PCR products amplified from internal transcribed spacer regions
with universal primers). Combined antifungal therapy with oral terbinafine and topical cyclopiroxolamide resulted in complete
remission of the fungal lesion within 1 month. Since 1944, only 29 cases of human M. praecox infection have been reported in the literature. The clinical features and treatment of these cases are reviewed. The prevalence
of M. praecox infection is probably underestimated, and systematic molecular identification could improve our understanding of the epidemiology
of this fungal dermatosis. 相似文献
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Nicolas Guigue Samuel Lardeux Alexandre Alanio Samia Hamane Marc Tabouret Stéphane Bretagne 《PloS one》2015,10(4)
Background
The Platelia Aspergillus Ag assay (Bio-Rad) is designed for detecting Aspergillus galactomannan (GM) and is widely used for diagnosing invasive aspergillosis but is hampered by variable occurrences of unreproducible positive results. Frequency and origin of these unreproducible results have not been formally studied.Methods
Different technicians simultaneously performed four tests on 550 consecutive sera from adult patients (Test#1-Test#2 for extraction#1 and Test#3-Test#4 for extraction#2). The samples were classified as confirmed negative [all tests with GM optical density index (GM-ODI) <0.5], confirmed positive (all tests with GM-ODI ≥0.5), extraction unreproducible positive (Test#1 and Test#2 ODIs ≥0.5, and Test#3 and Test#4 GM-ODIs <0.5, or conversely), and ELISA unreproducible positive (only one test with GM-ODI ≥0.5). The samples with positive and negative GM-ODIs within the assay coefficient of variation values were classified as non-conclusive. Four similar additional tests were performed after ≤72h storage at 4°C and a new GM test after 8 months at -20°C.Results
Five-hundred-twenty sera (94.5%) were confirmed negative, 15 (2.7%) confirmed positive, 4 (0.7%) extraction unreproducible positive, 6 (1.1%) ELISA unreproducible positive, and 5 (0.9%) non-conclusive. Upon retesting, the unreproducible positive results turned negative except for one which turned non-conclusive. The confirmed positive and non-conclusive had similar GM-ODIs (p>0.4) upon retesting after storage ≤72h at 4°C (n = 20) or eight months at -20°C (n = 17).Conclusions
Operational unreproducible positives represent 33% of the GM-positive results and a second sample evaluation appears mandatory to avoid useless investigations or treatments. When operational artifacts are excluded, GM remains stable at standard storage conditions. 相似文献7.
Dellière Sarah Gits-Muselli Maud Bretagne Stéphane Alanio Alexandre 《Mycopathologia》2020,185(5):783-800
Mycopathologia - Pneumocystis jirovecii pneumonia (PCP) is an important cause of morbidity in immunocompromised patients, with a higher mortality in non-HIV than in HIV patients. P. jirovecii is... 相似文献
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Alexandre Alanio Catherine Cordonnier Stéphane Bretagne 《Current fungal infection reports》2011,5(3):168-178
Azole resistance in Aspergillus fumigatus isolates is increasingly reported in different nosologic contexts with variable prevalence in different countries. Mutations
in the target of triazoles are widely described in azole-resistant clinical isolates. The recovery of mutated/resistant isolates
is described either in patients undergoing long-term azole treatment or after inhalation of environmentally acquired resistant
isolates. Acquisition in patients during azole therapy highlights the capacity of this fungus to adapt to its environment,
but it has a low impact in terms of public health, as interhuman transmission of A. fumigatus is uncommon. Environmentally acquired resistant isolates may propagate and affect populations at risk. The use of triazoles
as first-line therapy or prophylaxis could lead to selection of resistant isolates in patients, because most isolates harbor
azole cross-resistance. Although mold-active triazoles have provided major progress in the prophylaxis and treatment of Aspergillus infection, the increase of azole resistance could question their use in humans. 相似文献
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