Detection and species identification of microsporidial infections using SYBR Green real-time PCR


Polley, SD; Boadi, S; Watson, J; Curry, A; Chiodini, PL; (2011) Detection and species identification of microsporidial infections using SYBR Green real-time PCR. Journal of medical microbiology, 60 (4). pp. 459-466. ISSN 0022-2615 DOI: https://doi.org/10.1099/jmm.0.026781-0

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Abstract

Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron microscopy. This study describes a single SYBR Green real-time PCR assay for the simultaneous detection and species identification of such infections. This assay was highly sensitive, routinely detecting infections containing 400 parasites (g stool sample)(-1), whilst species identification was achieved by differential melt curves on a Corbett Life Science Rotor-Gene 3000. A modification of the QlAamp DNA tissue extraction protocol allowed the semi-automated extraction of DNA from stools for the routine diagnosis of microsporidial infection by real-time PCR. Of 168 stool samples routinely analysed for microsporidian spores, only five were positive by microscopy. By comparison, 17 were positive for microsporidial DNA by real-time analysis, comprising 14 Enterocytozoon bieneusi, one Encephalitozoon cuniculi and two separate Pleistophora species infections.

Item Type: Article
Keywords: POLYMERASE-CHAIN-REACTION, ENTEROCYTOZOON-BIENEUSI MICROSPORIDIA, SUBUNIT RIBOSOMAL-RNA, ENCEPHALITOZOON-CUNICULI, STOOL SPECIMENS, GIARDIA-LAMBLIA, ENTAMOEBA-HISTOLYTICA, IN-VITRO, IMMUNOCOMPROMISED, PATIENTS, QUANTITATIVE DETECTION
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Clinical Research
PubMed ID: 21183599
Web of Science ID: 289589300008
URI: http://researchonline.lshtm.ac.uk/id/eprint/848

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