PMID: 11899029Mar 20, 2002Paper

Development of a nested polymerase chain reaction assay for the detection and identification of Pythium insidiosum

Journal of Veterinary Internal Medicine
A M Grooters, Melaney K Gee

Abstract

Pythium insidiosum is an important cause of cutaneous and gastrointestinal disease in horses and dogs in the southeastern United States. Culture-based diagnosis of pythiosis is rarely definitive because production and identification of reproductive structures is difficult. The purpose of this study was to develop a polymerase chain reaction (PCR)-based assay for the identification of P insidiosum. Genomic DNA was extracted from 3 clinical isolates of P insidiosum and I isolate each of Pythium graminicola and Pythium arrhenomanes. The ITS I region of the ribosomal RNA gene of each isolate was amplified and sequenced, and the resultant sequences were aligned with published sequences for Pythium aphanidermatum, P acanthicum, and P myriotylum. A pair of P insidiosum-specific primers (PI-1 and PI-2) were designed from variable regions within the ITSI region. A nested PCR assay was developed in which the 1st round amplified the ITSI region by use of universal fungal primers. Second-round amplification utilized the internal P insidiosum-specific primers PI-1 and PI-2. Specificity of the assay was tested with DNA extracted from cultures of the following: 10 clinical isolates of P insidiosum and 1 isolate each of P graminicola, P irregu...Continue Reading

Citations

Jul 30, 2003·Mycological Research·Andrew M SchurkoGlen R Klassen
Sep 17, 2002·Veterinary Dermatology·Nadine R ZnajdaRosanna Marsella
Aug 21, 2012·Cornea·Onsiri ThanathaneeOlan Suwan-apichon
Jun 9, 2004·Journal of Clinical Microbiology·Andrew M SchurkoGlen R Klassen
Sep 15, 2004·Journal of Clinical Microbiology·Nongnuch VanittanakomNapaporn Tananuvat
Mar 18, 2009·Antimicrobial Agents and Chemotherapy·Ayrton S CavalheiroJanio M Santurio
May 4, 2011·Antimicrobial Agents and Chemotherapy·Erico Silva LoretoJanio Morais Santurio
Apr 18, 2012·Antimicrobial Agents and Chemotherapy·Deise Luiza MahlJanio Morais Santurio
Aug 25, 2015·Journal of Medical Microbiology·Angsana KeeratijarutTheerapong Krajaejun
Aug 27, 2009·Veterinary Dermatology·Rafael Torres NetoEduardo Bagagli
Jan 1, 2014·Veterinary Dermatology·William OldenhoffLauren Trepanier
Mar 6, 2007·The Veterinary Clinics of North America. Small Animal Practice·Sharon M Dial
Jun 16, 2015·JFMS Open Reports·Stuart WaltonVanessa R Barrs
Aug 21, 2013·Mycopathologia·Yordhathai ThongsriChularut Prariyachatigul
Dec 29, 2010·Current Microbiology·Sonia A BottonJanio Morais Santurio
Oct 8, 2003·Journal of Veterinary Internal Medicine·Amy M GrootersRandall C Thomas
Nov 5, 2015·Journal of the American Animal Hospital Association·David Lee DycusRyan Butler
Aug 13, 2003·The Veterinary Clinics of North America. Small Animal Practice·Amy M Grooters

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