2020
DOI: 10.3390/pathogens9080629
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Primary Amebic Meningoencephalitis Related to Groundwater in Costa Rica: Diagnostic Confirmation of Three Cases and Environmental Investigation

Abstract: During the first trimester of 2020, the Ministry of Health of Costa Rica reported the first three cases of primary amebic meningoencephalitis (PAM). In two cases, laboratory personnel of the hospitals preliminarily identified amoeboid forms in cerebrospinal fluid (CSF) samples. For the molecular confirmation of species, CSF samples were sent to our laboratory. We carried out microscopic analyses and exflagellation assays. Besides, samples were cultured in 2% casein hydrolysate medium and in non-nutrient agar p… Show more

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Cited by 4 publications
(4 citation statements)
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“…To confirm this result, another species-specific PCR using primers NfITS and applying serial dilutions to the DNA extracted from the EV fraction was performed, resulting in amplification until employing a DNA concentration of 3.125 ng, the last dilution in which the specific conventional PCR for N. fowleri resulted positive ( Figures 7C , D ). These results were confirmed with a N. fowleri species-specific qPCR ( Retana Moreira et al, 2020a , b ), in which mean CT = 30 for the dilution that contained 0.4 ng of DNA, mean CT = 31.5 for the dilution that contained 0.2 ng of DNA and mean CT = 32 for the dilution that contained 0.1 ng DNA.…”
Section: Resultssupporting
confidence: 60%
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“…To confirm this result, another species-specific PCR using primers NfITS and applying serial dilutions to the DNA extracted from the EV fraction was performed, resulting in amplification until employing a DNA concentration of 3.125 ng, the last dilution in which the specific conventional PCR for N. fowleri resulted positive ( Figures 7C , D ). These results were confirmed with a N. fowleri species-specific qPCR ( Retana Moreira et al, 2020a , b ), in which mean CT = 30 for the dilution that contained 0.4 ng of DNA, mean CT = 31.5 for the dilution that contained 0.2 ng of DNA and mean CT = 32 for the dilution that contained 0.1 ng DNA.…”
Section: Resultssupporting
confidence: 60%
“…Trophozoites of two clinic isolates of Naegleria fowleri from Costa Rica (accession numbers MT090627 and MT210902) ( Retana Moreira et al, 2020b ) were cultured in 75 cm 2 Nunc EasYFlask cell culture flasks (Thermo Fisher Scientific, Waltham, Massachusetts, United States) with 2% casein hydrolysate (Sigma Aldrich, Missouri, United States) culture medium, supplemented with 10% inactivated fetal bovine serum (Gibco, GranIsland, New York, United States) and antibiotics (penicillin/streptomycin). The flasks were incubated at 37°C, with daily observation of the cultures under an inverted microscope.…”
Section: Methodsmentioning
confidence: 99%
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“…Trophozoites of a clinical isolate of Naegleria fowleri from Costa Rica (accession number: MT090627) [ 34 ] were cultured in 75 cm 2 and 125 cm 2 flasks (Nunc EasYFlask ® , Thermo Fisher Scientific, Waltham, MA, USA) with 2% casein hydrolysate (Sigma Aldrich, St. Louis, MI, USA) culture medium, supplemented with 10% inactivated fetal bovine serum (Gibco, Grand Island, NY, USA) and antibiotics (penicillin/streptomycin). The flasks were incubated at 37 °C, with daily observation under an inverted microscope.…”
Section: Methodsmentioning
confidence: 99%