Identification of Echinococcus granulosus Genotypes G1 and G3 by SNPs Genotyping Assays

( ) is the causative agent of cystic echinococcosis in animals and humans. Different genotypes exhibit great diversity in their life cycle, host selectivity and pathogenicity. For this reason, the study of genetic variation within species is of importance for their epidemiological implication. We em...

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Bibliographic Details
Published in:Pathogens (Basel) 2021-01, Vol.10 (2), p.125
Main Authors: Bonelli, Piero, Dei Giudici, Silvia, Peruzzu, Angela, Mura, Lorena, Santucciu, Cinzia, Maestrale, Caterina, Masala, Giovanna
Format: Article
Language:English
Subjects:
Animal species
Assaying
Cysts
Deoxyribonucleic acid
Design
DNA
DNA sequencing
Echinococcosis
Echinococcus granulosus
Echinococcus granulosus sensu stricto
Epidemiology
Genetic diversity
Genetic testing
Genomes
Genotype & phenotype
Genotypes
genotypes G1 and G3
Genotyping
Identification
Life cycles
Melt temperature
Mitochondria
Nad5 gene
Pathogenicity
Pathogens
Polymerase chain reaction
Real time
real time PCR
Selectivity
Sheep
single nucleotide polymorphisms
Single-nucleotide polymorphism
Software
Zoonoses
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Summary:( ) is the causative agent of cystic echinococcosis in animals and humans. Different genotypes exhibit great diversity in their life cycle, host selectivity and pathogenicity. For this reason, the study of genetic variation within species is of importance for their epidemiological implication. We employed two SNP genotyping technologies to distinguish G1 and G3 ). genotypes. The genotypes of DNA samples (n = 28) extracted from hydatid cysts of different animal species were identified by amplification and sequencing of a fragment of the mitochondrial nad5 gene. Two SYBR green and three TaqMan real time PCR assays were developed for targeting of three nad5 informative positions (SNP758, 1123, and 1380) known to be able to discriminate G1 from G3. Genotyping by SYBR Green PCR based on cycle threshold (Ct) with melting temperature (Tm) analysis and performed on SNP1123 and SNP1380 failed to identify one DNA sample. TaqMan assays for SNP758, 1123 and 1380 effectively confirmed genotype identification obtained by Sanger sequencing. Our results demonstrated that the combination of the three Taqman assays developed in this study represents a valuable and cost effective tool alternative to DNA sequencing for genotyping.
ISSN:2076-0817
2076-0817
DOI:10.3390/pathogens10020125