. The purpose of this study is to optimize the DNA-based methods for species-specific diagnoses of microsporidiosis. The first specific aim will be to define the optimal methods for sample preparation, extraction of microsporidian DNA, and amplification of DNA by PCR for improving consistency in obtaining microsporidian DNA from clinical specimens (primarily stool and urine.) The second specific aim will be to define the levels of sensitivity, specificity, and reliability of methods used for detection of PCR DNA products. The detection methods that will be used include, ethidium bromide staining of PCR products, restriction fragment length polymorphism (PCR-RFLP), double-stranded DNA heteroduplex mobility shift analysis, and Southern analysis. The P.I. believes that the DNA-based methods incorporating the polymerase chain reaction (PCR) should result in more sensitive, specific and reliable levels of diagnosis than the histochemistry methods now being used, and will be important in epidemiology for defining the source and spread of microsporidia infections as ways to reduce exposure or prevent infection. She adds that species identification will be important for therapeutics because different species of microsporidia express different levels of sensitivity to some drugs (eg. albendazole) now being used to treat microsporidiosis.

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National Institute of Allergy and Infectious Diseases (NIAID)
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AIDS and Related Research Study Section 5 (ARRE)
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Tulane University
Internal Medicine/Medicine
Schools of Public Health
New Orleans
United States
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Didier, Elizabeth S; Bowers, Lisa C; Martin, Aaron D et al. (2010) Reactive nitrogen and oxygen species, and iron sequestration contribute to macrophage-mediated control of Encephalitozoon cuniculi (Phylum Microsporidia) infection in vitro and in vivo. Microbes Infect 12:1244-51
Nkinin, Stephenson W; Asonganyi, Tazoacha; Didier, Elizabeth S et al. (2007) Microsporidian infection is prevalent in healthy people in Cameroon. J Clin Microbiol 45:2841-6
Childs-Sanford, S E; Garner, M M; Raymond, J T et al. (2006) Disseminated microsporidiosis due to Encephalitozoon hellem in an Egyptian fruit bat (Rousettus aegyptiacus). J Comp Pathol 134:370-3
Didier, Elizabeth S; Weiss, Louis M (2006) Microsporidiosis: current status. Curr Opin Infect Dis 19:485-92
Juan-Salles, C; Garner, M M; Didier, E S et al. (2006) Disseminated encephalitozoonosis in captive, juvenile, cotton-top (Saguinus oedipus) and neonatal emperor (Saguinus imperator) tamarins in North America. Vet Pathol 43:438-46
Pandrea, Ivona; Mittleider, Derek; Brindley, Paul J et al. (2005) Phylogenetic relationships of methionine aminopeptidase 2 among Encephalitozoon species and genotypes of microsporidia. Mol Biochem Parasitol 140:141-52
Didier, Elizabeth S; Maddry, Joseph A; Brindley, Paul J et al. (2005) Therapeutic strategies for human microsporidia infections. Expert Rev Anti Infect Ther 3:419-34
Didier, Elizabeth S (2005) Microsporidiosis: an emerging and opportunistic infection in humans and animals. Acta Trop 94:61-76
Green, Linda C; Didier, Peter J; Bowers, Lisa C et al. (2004) Natural and experimental infection of immunocompromised rhesus macaques (Macaca mulatta) with the microsporidian Enterocytozoon bieneusi genotype D. Microbes Infect 6:996-1002
Dascomb, K; Frazer, T; Clark, R A et al. (2000) Microsporidiosis and HIV. J Acquir Immune Defic Syndr 24:290-2

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