Abstract
Blastocystis is a common food- and water-borne intestinal protist parasite of humans and many other animals. Blastocystis comprises multiple subtypes (STs) based on variability within the small subunit ribosomal (SSU rRNA) RNA gene. Though full-length reference sequences of the SSU rRNA gene are a current requirement to name a novel Blastocystis subtype, full-length reference sequences are not currently available for all subtypes. In the present study, Oxford Nanopore MinION long-read sequencing was employed to generate full-length SSU rRNA sequences for seven new Blastocystis subtypes for which no full-length references currently exist: ST21, ST23, ST24, ST25, ST26, ST27, and ST28. Phylogenetic analyses and pairwise distance matrixes were used to compare full-length and partial sequences of the two regions that are most commonly used for subtyping. Analyses included Blastocystis nucleotide sequences obtained in this study (ST21 and ST23–ST28) and existing subtypes for which full-length reference sequences were available (ST1–ST17 and ST29). The relationships and sequence variance between new and existing subtypes observed in analyses of different portions of the SSU rRNA gene are discussed. The full-length SSU rRNA reference sequences generated in this study provide essential new data to study and understand the relationships between the genetic complexity of Blastocystis and its host specificity, pathogenicity, and epidemiology.
Highlights
Blastocystis sp. is one of the most common intestinal parasites observed in humans and has a global distribution [1,2]
The five samples included in this study were selected based on the results of screening via MiSeq sequencing of an approximately 500 bp region of the SSU rRNA gene and were found to contain ST21, ST23, ST24, ST25, ST26, ST27, and/or ST28
Fulllength sequences of the SSU rRNA gene allow for the construction of phylogenies that can aide in more accurately describing the relationship between Blastocystis subtypes from humans and other animal hosts
Summary
Blastocystis sp. is one of the most common intestinal parasites observed in humans and has a global distribution [1,2]. Is one of the most common intestinal parasites observed in humans and has a global distribution [1,2] It is spread via the fecal–oral route, with contaminated food and water being likely means of transmission [3,4,5]. One factor thought to be key in explaining the pathogenicity and zoonotic potential of Blastocystis is the existence of a remarkable degree of genetic variability within the genus. This genetic variability has far been largely described using variability within the small subunit ribosomal RNA gene (SSU rRNA gene). Subtype designations are assigned using a numbering scheme that is chronological and based on publication order [9]
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