Abstract
Granulomatous amoebic encephalitis (GAE) is a usually fatal disease caused by the free-living amoebae Balamuthia mandrillaris and Acanthamoeba spp. The intent of this study was to determine if the polymerase chain reaction (PCR) could be used retrospectively to detect amoeba mitochondrial 16S ribosomal ribonucleic acid gene deoxyribonucleic acid (DNA) in confirmed archival tissue sections from GAE cases stored in our laboratories for 1 to 34 years. The DNA was extracted from deparaffinized sections, and appropriate primer sets for each of the two amoebae were used for DNA detection. Indirect immunofluorescent staining (IIF) of tissue sections was used as the standard for identification of amoebae against which the PCR results were compared. Sixty slides from a total of 56 cases were processed by PCR for amoeba 16S DNA. In 28 (47%) slides, there was agreement between the IIF and PCR results. In 41 of the slides (52%), no DNA was detected after PCR. In one slide (1%), the PCR and IIF results did not agree. While PCR supported IIF findings in about half of the slides, there are significant limitations in amoeba DNA identifications in formalin-fixed brain tissues. Degradation of amoeba DNA because of formalin fixation was probably a factor in limiting valid results.
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Acknowledgments
The research was supported in part by a grant from the Emerging Infectious Diseases Program of the Centers for Disease Control and Prevention (U50/CCU915548-09) to Dr. Carol Glaser of the California Department of Public Health, Richmond, whom the authors thank for her encouragement and support. We thank Rama Sriram (CDC) for her technical expertise and assistance. The research performed in this study was in compliance with the laws of the USA and the authors’ respective institutions.
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Yagi, S., Schuster, F.L. & Visvesvara, G.S. Demonstration of Balamuthia and Acanthamoeba mitochondrial DNA in sectioned archival brain and other tissues by the polymerase chain reaction. Parasitol Res 102, 211–217 (2008). https://doi.org/10.1007/s00436-007-0749-7
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DOI: https://doi.org/10.1007/s00436-007-0749-7