Visceral leishmaniasis in acute myeloid leukemia revealed on peripheral blood smear

International audienceKey Clinical Message Images of parasitic forms of Leishmania infantum are typical in the hands of a skilled expert but should be known by biologists of Hematology Department. In an endemic region, the diagnosis of visceral leishmaniasis (VL) must be considered because of its potential role in accelerating hematological malignancy

Evaluation of Two Commercial Kits on the Automated ELITe InGenius PCR Platform for Molecular Diagnosis of Toxoplasmosis

International audienceMolecular diagnosis of toxoplasmosis is essential for establishing the diagnosis of congenital contaminations and for primary infection or reactivation of immunocompromised patients. An integrated extraction and real-time PCR-based system is of particular interest in this context. Commercial kits for automated extraction and amplification steps are now available. Herein, we assessed two commercial PCR assays for this diagnosis, those of Bio-Evolution and Elitech, on the ELITe InGenius platform. The Bio-Evolution assay showed a specificity and a sensitivity of 100% on clinical samples, but a lower analytical detection threshold than the Elitech assay. The latter showed a specificity of 100% and a sensitivity of 96%. The SP1000 cartridges, which allow DNA extraction from 1 mL of template, showed interesting performances on amniotic fluid samples. Overall, the two kits had good performances on the InGenius platform, which offers a turn-key solution suitable for the molecular diagnosis of toxoplasmosis

Enterocytozoon bieneusi, a human pathogen

Although brought to the forefront in the 1980s with the AIDS pandemic, microsporidia infecting humans are still little known. Enterocytozoon bieneusi, by far the most frequent microsporidia species causing diseases in humans, is responsible for intestinal illness in both non- and immunocompromised patients. This species presents an astonishing genetic diversity with more than 500 genotypes described, some of which have a strong zoonotic potential. Indeed, E. bieneusi infects a broad array of hosts, from wild to domestic animals. This emerging eukaryotic pathogen has thus been associated with foodborne/waterborne outbreaks. Several molecular assays have been developed to enhance its diagnosis or for epidemiological purposes, providing valuable new data. Here, we propose an overview of the current knowledge on this major species among the microsporidia, so far rather neglected in human medicine

Current status of intestinal parasitosis and microsporidiosis in industrialized countries: Results from a prospective study in France and Luxembourg.

BackgroundHuman intestinal parasitosis and microsporidiosis are a global health concern, mostly in endemic areas but should not be neglected elsewhere. Recent nationwide epidemiological data are scarce, especially from primary health care and developed countries. Diagnosis by molecular tools are increasing and several commercial gastrointestinal panel assays including protozoans and/or helminths are now available. These news tools improve the knowledge into real human parasite epidemiology. This study provides an epidemiological update on intestinal parasites found in primary health care in France and Luxembourg.Methodology/principal findingsTwo thousand fifty-six stools from primary health care patients were analyzed for the presence of intestinal parasites (IPs) during two different seasons of 2022, the winter and the summer, corresponding to more than 1500 patients from all over France and Luxembourg. Parasite detection was performed combining standard microscopy (merthiolate-iodine-formaldehyde and Bailenger concentration procedures) with two molecular panel assays (AMPLIQUICK Fecal Pretreatment, AMPLIQUICK Protozoans and AMPLIQUICK Helminths, BIOSYNEX, France). The prevalence of IPs in primary care patients reached 33.2%. Blastocystis sp. and Dientamoeba fragilis were the most frequently detected parasites in 20.5% and 13.1% of patients, respectively. Coinfection with two or more parasites was detected in 9.9% of patients. For some parasites, patterns according to gender, age, geography or season have been observed.Conclusion/significanceThe high prevalence of pathogenic IPs (about 7%) underlines the importance of investigating gastrointestinal disorders through parasite examination, even in developed countries. The detection of parasites, pathogenic or not, remains a marker of the faecal-oral route of transmission and results should be interpreted accordingly. Parasites molecular characterization give new insights and should encourage further research as industrialized countries are not exempt of parasitic circulation and a better survey is necessary

Malignant Aspergillus flavus Otitis Externa with Jugular Thrombosis

We report a case of malignant otitis externa with jugular vein thrombosis caused by Aspergillus flavus. Magnetic resonance imaging revealed an unusual ink smudge pattern deep in a cervical abscess. The pattern was consistent with mycetoma and may be important for diagnosing these life-threatening infections

Multicenter comparative study of Enterocytozoon bieneusi DNA extraction methods from stool samples, and mechanical pretreatment protocols evaluation

International audienceNowadays, the use of qPCR for the diagnosis of intestinal microsporidiosis is increasing. There are several studies on the evaluation of qPCR performance but very few focus on the stool pretreatment step before DNA extraction, which is nevertheless a crucial step. This study focuses on the mechanical pretreatment of stools for Enterocytozoon bieneusi spores DNA extraction. Firstly, a multicenter comparative study was conducted evaluating seven extraction methods (manual or automated) including various mechanical pretreatment. Secondly, several durations and grinding speeds and types of beads were tested in order to optimize mechanical pretreatment. Extraction methods of the various centers had widely-varying performances especially for samples with low microsporidia loads. Nuclisens® easyMAG (BioMérieux) and Quick DNA Fecal/Soil Microbe Microprep kit (ZymoResearch) presented the best performances (highest frequencies of detection of low spore concentrations and lowest Ct values). Optimal performances of mechanical pretreatment were obtained by applying a speed of 30 Hz during 60 s with the TissueLyser II (Qiagen) using commercial beads of various materials and sizes (from ZymoResearch or MP Biomedicals). Overall, the optimal DNA extraction method for E. bieneusi spores contained in stool samples was obtained with a strong but short bead beating using small-sized beads from various materials

Comparison of DNA extraction methods and real-time PCR assays for the detection of blastocystis sp. in stool specimens

International audienceDiagnosis of Blastocystis in stool may be challenging, as microscopic examination and culture-based methods have demonstrated low sensitivity. Molecular detection assays are now available for this enteric parasite, based on “in-house” or commercial-developed techniques. The aim of this study was to assess and compare the performance of (i) two DNA extraction methods (manual versus automated), and (ii) four qPCR assays (three “in-house” and one commercialized), for detection of Blastocystis sp. in human stools. One hundred and forty stools were included, among which 76 were confirmed to be positive for Blastocystis. The manual DNA extraction method allowed for the identification of significantly more positive specimens than the automated method (p < 0.05). In particular, specimens with a low parasite load were negative when DNA was extracted with the automated process. The four qPCR assays also had variable performances, with the commercialized assay being the most sensitive (84%) but the least specific (82%). Overall, for all qPCR assays, the specificity decreased when the sensitivity increased. Blastocystis’ subtype, notably the subtype 4, influenced these performances. Our results indicate that the positivity rate for the detection of Blastocystis in stools could be variable according to the DNA extraction method and the qPCR assay used. These pitfalls need to be considered for the selection of method and interpretation of results, particularly considering the search of this intestinal parasite in a donor before fecal microbiota transplantation

Comparison of DNA Extraction Methods and Real-Time PCR Assays for the Detection of Blastocystis sp. in Stool Specimens

Diagnosis of Blastocystis in stool may be challenging, as microscopic examination and culture-based methods have demonstrated low sensitivity. Molecular detection assays are now available for this enteric parasite, based on “in-house” or commercial-developed techniques. The aim of this study was to assess and compare the performance of (i) two DNA extraction methods (manual versus automated), and (ii) four qPCR assays (three “in-house” and one commercialized), for detection of Blastocystis sp. in human stools. One hundred and forty stools were included, among which 76 were confirmed to be positive for Blastocystis. The manual DNA extraction method allowed for the identification of significantly more positive specimens than the automated method (p &lt; 0.05). In particular, specimens with a low parasite load were negative when DNA was extracted with the automated process. The four qPCR assays also had variable performances, with the commercialized assay being the most sensitive (84%) but the least specific (82%). Overall, for all qPCR assays, the specificity decreased when the sensitivity increased. Blastocystis’ subtype, notably the subtype 4, influenced these performances. Our results indicate that the positivity rate for the detection of Blastocystis in stools could be variable according to the DNA extraction method and the qPCR assay used. These pitfalls need to be considered for the selection of method and interpretation of results, particularly considering the search of this intestinal parasite in a donor before fecal microbiota transplantation.</jats:p