@article{mbs:/content/journal/jmm/10.1099/jmm.0.026781-0, author = "Polley, Spencer D. and Boadi, Samuel and Watson, Julie and Curry, Alan and Chiodini, Peter L.", title = "Detection and species identification of microsporidial infections using SYBR Green real-time PCR", journal= "Journal of Medical Microbiology", year = "2011", volume = "60", number = "4", pages = "459-466", doi = "https://doi.org/10.1099/jmm.0.026781-0", url = "https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.026781-0", publisher = "Microbiology Society", issn = "1473-5644", type = "Journal Article", keywords = "CI, confidence limit", keywords = "Ct, cycle threshold", keywords = "SSU, small subunit", keywords = "TEM, transmission electron microscopy", 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 QIAamp 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.", }