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Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients.
J Clin Microbiol. 1997 Mar; 35(3):652-5.JC

Abstract

A routine assay based on the PCR was developed for the detection of Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal samples. Two oligonucleotide primer pairs from a conserved region in the small-subunit rRNA genes of E. bieneusi (primer pair V1 and EB450) and E. intestinalis (primer pair V1 and SI500) were used to amplify microsporidian DNA. We achieved specific amplification of a 382-bp DNA fragment in E. intestinalis and a 353-bp DNA fragment in E. bieneusi. Boiling of the samples appeared to be most effective for DNA extraction. Fecal samples containing fewer than 10 microsporidia gave a positive result in the PCR assay. Fecal specimens from 30 human immunodeficiency virus-infected patients with microsporidiosis and fecal specimens from 42 patients suspected of having microsporidiosis were investigated by the PCR assay. The PCR assay was validated against standard staining methods (the Uvitex 2B and Chromotrope 2R staining methods) and immunofluorescence assay specific for E. intestinalis. This comparative study has shown that PCR improved species determination and can thus be considered a fast and reliable method for the detection and identification of each intestinal species.

Authors+Show Affiliations

Centre Hospitalo-Universitaire de la Pitié-Salpêtrière, Unité INSERM 313, Paris, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9041406

Citation

Ombrouck, C, et al. "Specific PCR Assay for Direct Detection of Intestinal Microsporidia Enterocytozoon Bieneusi and Encephalitozoon Intestinalis in Fecal Specimens From Human Immunodeficiency Virus-infected Patients." Journal of Clinical Microbiology, vol. 35, no. 3, 1997, pp. 652-5.
Ombrouck C, Ciceron L, Biligui S, et al. Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients. J Clin Microbiol. 1997;35(3):652-5.
Ombrouck, C., Ciceron, L., Biligui, S., Brown, S., Marechal, P., van Gool, T., Datry, A., Danis, M., & Desportes-Livage, I. (1997). Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients. Journal of Clinical Microbiology, 35(3), 652-5.
Ombrouck C, et al. Specific PCR Assay for Direct Detection of Intestinal Microsporidia Enterocytozoon Bieneusi and Encephalitozoon Intestinalis in Fecal Specimens From Human Immunodeficiency Virus-infected Patients. J Clin Microbiol. 1997;35(3):652-5. PubMed PMID: 9041406.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients. AU - Ombrouck,C, AU - Ciceron,L, AU - Biligui,S, AU - Brown,S, AU - Marechal,P, AU - van Gool,T, AU - Datry,A, AU - Danis,M, AU - Desportes-Livage,I, PY - 1997/3/1/pubmed PY - 1997/3/1/medline PY - 1997/3/1/entrez SP - 652 EP - 5 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 35 IS - 3 N2 - A routine assay based on the PCR was developed for the detection of Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal samples. Two oligonucleotide primer pairs from a conserved region in the small-subunit rRNA genes of E. bieneusi (primer pair V1 and EB450) and E. intestinalis (primer pair V1 and SI500) were used to amplify microsporidian DNA. We achieved specific amplification of a 382-bp DNA fragment in E. intestinalis and a 353-bp DNA fragment in E. bieneusi. Boiling of the samples appeared to be most effective for DNA extraction. Fecal samples containing fewer than 10 microsporidia gave a positive result in the PCR assay. Fecal specimens from 30 human immunodeficiency virus-infected patients with microsporidiosis and fecal specimens from 42 patients suspected of having microsporidiosis were investigated by the PCR assay. The PCR assay was validated against standard staining methods (the Uvitex 2B and Chromotrope 2R staining methods) and immunofluorescence assay specific for E. intestinalis. This comparative study has shown that PCR improved species determination and can thus be considered a fast and reliable method for the detection and identification of each intestinal species. SN - 0095-1137 UR - https://www.unboundmedicine.com/medline/citation/9041406/Specific_PCR_assay_for_direct_detection_of_intestinal_microsporidia_Enterocytozoon_bieneusi_and_Encephalitozoon_intestinalis_in_fecal_specimens_from_human_immunodeficiency_virus_infected_patients_ L2 - https://journals.asm.org/doi/10.1128/jcm.35.3.652-655.1997?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -