Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients

被引:51
作者
Ombrouck, C
Ciceron, L
Biligui, S
Brown, S
Marechal, P
vanGool, T
Datry, A
Danis, M
DesportesLivage, I
机构
[1] CNRS,INST SCI VEGETALES,LAB CYTOMETRIE,F-91198 GIF SUR YVETTE,FRANCE
[2] MUSEUM NATL HIST NAT,LAB BIOL PARASITAIRE,F-75231 PARIS,FRANCE
[3] UNIV AMSTERDAM,ACAD MED CTR,DEPT MED MICROBIOL,NL-1105 AZ AMSTERDAM,NETHERLANDS
关键词
D O I
10.1128/JCM.35.3.652-655.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 S1500) 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.
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页码:652 / 655
页数:4
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