Comparative Genomic and Transcriptomic Analysis of Naegleria fowleri Clinical and Environmental Isolates

Abstract

N. fowleri , the only known Naegleria species to infect humans, causes fatal brain disease. PAM cases from 1965 to 2016 showed <20 cases per year globally.

The First Molecular Genotyping of Naegleria fowleri Causing Primary Amebic Meningoencephalitis in Thailand With Epidemiology and Clinical Case Reviews

“…However, the 12S rRNA and ITS1 sequences showed disagreement in the genotype between each other ( Figures 3 , 5 ). Interestingly, this discrepancy was also observed in the isolate V1005 from the PAM patient in Australia on both mitochondrial 12S rRNA and nuclear ITS1 genes ( Joseph et al, 2021 ). It was previously reported that genotyping based on 12S rRNA and ITS1 was consistent between the two loci ( Zhou et al., 2003 ).…”

Differential Growth Rates and In Vitro Drug Susceptibility to Currently Used Drugs for Multiple Isolates of Naegleria fowleri

“…In addition, we wanted to compare the drug susceptibilities of the clinical isolates with their genotypes. There is published internal transcribed spacer (ITS) genotype information, determined based upon the techniques described by Zhou et al, for all of the isolates except Nf69 ( 39 , 40 ). After obtaining Sanger sequencing results for the PCR products for Nf69, we trimmed off poor-quality base pairs and aligned the sequences obtained for the forward primer sequencing samples as well as the reverse primer sequencing samples to attain a 446-bp sequence.…”

“…A recent study by Joseph et al sequenced the genomes for each of the isolates tested in our study, and we downloaded and processed these data in order to perform the aforementioned genotyping technique and compare the genotype results attained with the system created by De Jonckheere with those reported based on the work of Zhou et al ( 39 – 41 ). Using Geneious Prime software, we pairwise aligned the forward and reverse reads for each clinical isolate and then mapped these to each of the 2 ITS primers.…”

“…To compare genotype determinations for each of the two methods that are reported in the literature ( 39 , 41 ), we accessed the genome sequences recently published under BioProject no. PRJNA642022 and imported them into Geneious Prime software as paired reads in order to obtain pairwise alignments (options: Illumina, paired end [inward pointing]; pairs of files, 500 bp) for each clinical isolate included in this study ( 40 ). We then mapped the reads to the ITSFWD and ITSREV primers using the Geneious mapper with medium-low sensitivity, allowing iterations up to 5 times for fine tuning (all other options were set to default).…”

The State of Art of Extracellular Traps in Protozoan Infections (Review)

“…Naegleria fowleri ( N. fowleri ) is a free-living amoeba that can infect humans through the nasal mucosa and cause central nervous system diseases, such as primary amoebic meningoencephalitis (PAM) ( 60 , 61 ). Because NETs may be an important factor in the fixing and killing of invading microorganisms, the study of the role of NETs in central nervous system diseases caused by N. fowleri will help us to better understand the relevant mechanisms and regulatory pathways of parasites involved in the occurrence and development of diseases.…”