Acanthamoeba Can Be Differentiated by the Polymerase Chain Reaction and Simple Plating Assays |
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Authors: | Naveed A Khan Edward L Jarroll Timothy A Paget |
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Institution: | (1) Department of Pediatric Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA, US;(2) Department of Biological Sciences, Hardy Building, University of Hull, Hull, HU6 7RX, England, UK, GB;(3) Department of Biology, Northeastern University, Boston, MA 02115, USA, US |
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Abstract: | Acanthamoeba are opportunistic pathogens with invasive and noninvasive species. For clinical purposes it is important to differentiate
potentially pathogenic from nonpathogenic isolates. For the rapid and sensitive identification of Acanthamoeba at the genus level, we used a polymerase chain reaction (PCR)-based method which detected as few as five cells. Further,
we tested nine isolates of Acanthamoeba for their ability to produce cytopathic effects (CPE) on corneal epithelial cells. On the basis of the results, Acanthamoeba were divided into pathogenic or nonpathogenic groups. However, because CPE assays are not available to every diagnostic laboratory,
we developed a simple plating assay based on osmotolerance which correlated well with the CPE assays. Pathogenic Acanthamoeba showed growth on higher osmolarity (agar plates containing one molar mannitol), while growth of nonpathogens was inhibited
on these plates. In conclusion, we have developed methods for the rapid identification and differentiation of Acanthamoeba.
Received: 5 January 2001/Accepted: 6 February 2001 |
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