Development and Evaluation of a Real-Time PCR Assay for Detection and Quantification of Blastocystis Parasites in Human Stool Samples: Prospective Study of Patients with Hematological Malignancies
| Autor: | Hicham El Alaoui, Philippe Poirier, Aurélie Albert, Frédéric Delbac, Ivan Wawrzyniak, Valérie Livrelli |
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| Přispěvatelé: | Laboratoire Microorganismes : Génome et Environnement (LMGE), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA), Service de Bactériologie [CHU Clermont-Ferrand], CHU Gabriel Montpied [Clermont-Ferrand], CHU Clermont-Ferrand-CHU Clermont-Ferrand, CHU Clermont-Ferrand, Laboratoire Microorganismes : Génome et Environnement - Clermont Auvergne (LMGE), Université Clermont Auvergne (UCA)-Centre National de la Recherche Scientifique (CNRS), Evolution des bactéries pathogènes et susceptibilité génétique de l'hôte (JE2526), Institut National de la Recherche Agronomique (INRA)-Université d'Auvergne - Clermont-Ferrand I (UdA), Service Bactériologie Mycologie Parasitologie |
| Rok vydání: | 2011 |
| Předmět: |
Male
Pathology Blastocystis Infections Polymerase Chain Reaction Parasite load law.invention Feces MESH: Blastocystis Infections 0302 clinical medicine [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases law Prospective Studies Polymerase chain reaction MESH: Aged 0303 health sciences MESH: Middle Aged MESH: DNA Ribosomal MESH: Molecular Typing MESH: Feces Middle Aged Subtyping 3. Good health Real-time polymerase chain reaction Hematologic Neoplasms Female [SDV.IB]Life Sciences [q-bio]/Bioengineering MESH: Blastocystis Adult Microbiology (medical) medicine.medical_specialty 030231 tropical medicine MESH: Parasitology MESH: DNA Protozoan Biology DNA Ribosomal Sensitivity and Specificity 03 medical and health sciences medicine Humans [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Aged Blastocystis MESH: Humans 030306 microbiology MESH: Polymerase Chain Reaction MESH: Adult DNA Protozoan Ribosomal RNA biology.organism_classification Virology MESH: Male MESH: Prospective Studies MESH: Sensitivity and Specificity Molecular Typing Parasitology RNA Ribosomal MESH: RNA Ribosomal MESH: Female MESH: Hematologic Neoplasms |
| Zdroj: | Journal of Clinical Microbiology Journal of Clinical Microbiology, 2011, 49 (3), pp.975-983. ⟨10.1128/JCM.01392-10⟩ Journal of Clinical Microbiology, American Society for Microbiology, 2011, 49 (3), pp.975-983. ⟨10.1128/JCM.01392-10⟩ |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.01392-10 |
| Popis: | Blastocystis anaerobic parasites are widespread worldwide in the digestive tract of many animal species, including humans. Epidemiological Blastocystis studies are often limited by the poor sensitivity of standard parasitological assays for its detection. This report presents a highly sensitive real-time quantitative PCR (qPCR) assay developed to detect Blastocystis parasites in stool samples. The assay targets a partial sequence of the Blastocystis small ribosomal subunit (SSU) rRNA gene, allowing subtyping (ST) of Blastocystis isolates by direct sequencing of qPCR products. This qPCR method was assessed in a prospective study of 186 patients belonging to two cohorts—a group of 94 immunocompromised patients presenting hematological malignancies and a control group of 92 nonimmunocompromised patients. Direct-light microscopy and xenic in vitro stool culture analysis showed only 29% and 52% sensitivity, respectively, compared to our qPCR assay. Of the 27 (14.5%) Blastocystis -positive patients, 8 (4%) experienced digestive symptoms. No correlation was found between symptomatic patients and immune status, parasite load, or parasite subtypes, although subtyping of all isolates revealed a high (63.0%) prevalence of ST4. Two unexpected avian subtypes were found, i.e., ST6 and ST7, which are frequently isolated in Asia but rarely present in Western countries. In conclusion, this qPCR proved by far the most sensitive of the tested methods and allowed subtype determination by direct sequencing of qPCR products. New diagnostic tools such as the qPCR are essential for evaluating the clinical relevance of Blastocystis subtypes and their role in acute or chronic digestive disorders. |
| Databáze: | OpenAIRE |
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