Towards a rapid sequencing-based molecular surveillance and mosaicism investigation of Toxoplasma gondii

Vilares, Anabela; Borges, Vítor; Sampaio, Daniel A.; Ferreira, Idalina; Martins, Susana; Vieira, Luı́s; Gargaté, Maria João; Gomes, João Paulo

doi:10.1007/s00436-019-06523-3

Cited by 4 publications

(3 citation statements)

References 47 publications

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“…The acquisition of nucleotide sequence data of Toxoplasma gondii is hard to achieve due to the difficulty of isolating the parasite from asymptomatic hosts; thus, researchers must attempt genotyping from clinical samples where the DNA of the parasite is heavily diluted within the host’s DNA [ 5 , 13 ]. To deepen the analysis of the mixed infections found in the SAG3 locus, we cloned and sequenced DNA from clinical samples that showed a triple infection pattern by PCR-RFLP.…”

Section: Discussionmentioning

confidence: 99%

“…Intragenic recombinant genotypes can be generated during the sexual reproduction of T. gondii . In addition, during the asexual reproduction cycle within intermediate hosts (and in nonintestinal epithelial cells of felines), single nucleotide polymorphisms (SNPs) may arise [ 4 , 5 ]. Regions with a tropical climate and a high density of definitive hosts are expected to have a high genetic diversity of the parasite and a lower probability of finding clonal genotypes/sequences [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

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SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean

et al. 2022

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Background Currently, more than 300 genotypes of Toxoplasma gondii (T. gondii) have been described throughout the world, demonstrating its wide genetic diversity. The SAG3 locus is one of the genes included in the genotyping panel of this parasite. It is associated with its virulence since it participates during the invasion process of the host cells. Therefore, cloning, sequencing, and bioinformatic analysis were used to deepen the understanding of the SAG3 locus genetic diversity of T. gondii in blood samples from feral cats. Results Six different SAG3 sequences were detected, five of which were detected in one feline. Three sequences were first reported here; one of them was an intragenic recombinant. In the cladogram, four out of ten SAG3 sequences did not share nodes with others reported worldwide. Conclusions Cloning and sequencing of samples with more than one restriction pattern by PCR-RFLP were very helpful tools to demonstrate the presence of more than three genotypes of T. gondii in the blood of feral cats from southeastern Mexico. This suggests a potential mixed infection of multiple T. gondii strains and high genetic diversity of the parasites in felines in this tropical region of Mexico.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean

et al. 2022

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“…Furthermore, whole genome sequencing (WGS) provides the most detailed information about genetic variability. However, WGS requires highly concentrated DNA and may not be suitable for laboratories with more limited resources [ 16 ]. In addition, WGS of T. gondii is a bioinformatically challenging task due to the size of its genome, approximately 65 Mb [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

Genotyping of European Toxoplasma gondii strains by a new high-resolution next-generation sequencing-based method

Joeres,

Maksimov,

Höper

et al. 2023

Eur J Clin Microbiol Infect Dis

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Purpose A new high-resolution next-generation sequencing (NGS)-based method was established to type closely related European type II Toxoplasma gondii strains. Methods T. gondii field isolates were collected from different parts of Europe and assessed by whole genome sequencing (WGS). In comparison to ME49 (a type II reference strain), highly polymorphic regions (HPRs) were identified, showing a considerable number of single nucleotide polymorphisms (SNPs). After confirmation by Sanger sequencing, 18 HPRs were used to design a primer panel for multiplex PCR to establish a multilocus Ion AmpliSeq typing method. Toxoplasma gondii isolates and T. gondii present in clinical samples were typed with the new method. The sensitivity of the method was tested with serially diluted reference DNA samples. Results Among type II specimens, the method could differentiate the same number of haplotypes as the reference standard, microsatellite (MS) typing. Passages of the same isolates and specimens originating from abortion outbreaks were identified as identical. In addition, seven different genotypes, two atypical and two recombinant specimens were clearly distinguished from each other by the method. Furthermore, almost all SNPs detected by the Ion AmpliSeq method corresponded to those expected based on WGS. By testing serially diluted DNA samples, the method exhibited a similar analytical sensitivity as MS typing. Conclusion The new method can distinguish different T. gondii genotypes and detect intra-genotype variability among European type II T. gondii strains. Furthermore, with WGS data additional target regions can be added to the method to potentially increase typing resolution.

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Genotyping of human isolates from human toxoplasmosis outbreak: Restriction Fragment Length Polymorphism and Sanger Sequencing

Fernandes,

Bräunig,

Machado

et al. 2024

Braz J Microbiol

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Towards a rapid sequencing-based molecular surveillance and mosaicism investigation of Toxoplasma gondii

Cited by 4 publications

References 47 publications

SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean

SAG3 Toxoplasma gondii cloning reveals unexpected fivefold infection in the blood of feral cats in the Mexican Caribbean

Genotyping of European Toxoplasma gondii strains by a new high-resolution next-generation sequencing-based method

Genotyping of human isolates from human toxoplasmosis outbreak: Restriction Fragment Length Polymorphism and Sanger Sequencing

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