Species-specific identification of microsporidia in stool and intestinal biopsy specimens by the polymerase chain reaction |
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| Authors: | N P Kock H Petersen T Fenner I Sobottka C Schmetz P Deplazes N J Pieniazek H Albrecht J Schottelius |
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| Institution: | (1) Section of Parasitology, Bernard Nocht Institute for Tropical Medicine, Bernard-Nocht-Straße 74, D-20359 Hamburg, Germany;(2) Institute for Clinical Pathology and Microbiology, Dres. Fenner and Partners, Hamburg, Germany;(3) Institute of Microbiology and Immunology, University Hospital Eppendorf, Hamburg, Germany;(4) Electron Microscopy Laboratory, Bernard Nocht Institute for Tropical Medicine, Bernard-Nocht-Straße 74, D-20359 Hamburg, Germany;(5) Institute of Parasitology, University of Zürich, Zürich, Switzerland;(6) Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA;(7) Department of Internal Medicine, University Hospital Eppendorf, Hamburg, Germany |
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| Abstract: | In view of the increasing number of cases of human microsporidiosis, simple and rapid methods for clear identification of microsporidian parasites to the species level are required. In the present study, the polymerase chain reaction (PCR) was used for speciesspecific detection ofEncephalitozoon cuniculi, Encephalitozoon hellem, Encephalitozoon (Septata) intestinalis, andEnterocytozoon bieneusi in both tissue and stool. Using stool specimens and intestinal biopsies of patients infected withEnterocytozoon bieneusi (n=9),Encephalitozoon spp. (n=2), andEncephalitozoon intestinalis (n=1) as well as stool spiked with spores ofEncephalitozoon cuniculi andEncephalitozoon hellem and tissue cultures ofEncephalitozoon cuniculi andEncephalitozoon hellem, three procedures were developed to produce PCR-ready DNA directly from the samples. Specific detection of microsporidian pathogens was achieved in the first PCR. The subsequent nested PCR permitted species determination and verified the first PCR products. Without exception, the PCR assay confirmed electron microscopic detection ofEnterocytozoon bieneusi andEncephalitozoon intestinalis in stool specimens and their corresponding biopsies and in spiked stool samples and tissue cultures infected withEncephalitozoon cuniculi andEncephalitozoon hellem. Moreover, identification ofEncephalitozoon spp. could be specified asEncephalitozoon intestinalis. Whereas standard methods such as light and transmission electron microscopy may lack sensitivity or require more time and special equipment, the PCR procedure described facilitates speciesspecific identification of microsporidian parasites in stool, biopsies, and, probably, other samples in about five hours. |
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