Detection and genetic characterization of Echinococcus granulosus mitochondrial DNA in serum and formalin-fixed paraffin embedded cyst tissue samples of cystic echinococcosis patients
Autoři:
Maryam Moradi aff001; Ahmad Reza Meamar aff002; Lame Akhlaghi aff002; Mona Roozbehani aff002; Elham Razmjou aff002
Působiště autorů:
Department of Parasitology and Mycology, School of Medicine, International Campus, Iran University of Medical Sciences, Tehran, Iran
aff001; Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
aff002
Vyšlo v časopise:
PLoS ONE 14(10)
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pone.0224501
Souhrn
Cystic echinococcosis (CE) is a worldwide zoonotic disease caused by the larval stage of Echinococcus granulosus. We investigated the presence of E. granulosus-specific DNA in the serum of CE patients by detecting the cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) mitochondrial genes. Serum and formalin-fixed paraffin embedded (FFPE) cyst tissue samples of 80 CE patients were analyzed. The extracted DNA of samples was submitted to PCR amplification of cox1 and nad1 genes, and products were sequenced and genotyped. Nineteen (23.8%; 95% CI 15.8–34.1) serum and 78 (97.5%; 95% CI 91.3–99.3) FFPE cyst tissue samples were successfully amplified with at least one gene. Echinococcus DNA was detected in the sera of 15.0% (95% CI: 8.8–24.4) and 10.0% (95% CI: 5.2–18.5) and in cyst tissue of 91.3% (95% CI: 83.0–95.7) and 83.8% (95% CI: 74.2–90.3) of 80 patients by cox1 and nad1 gene, respectively. Four genotypes of E. granulosus were distinguished in the CE patients, with predominance of genotype G1, followed by G3, G2, and G6. The finding of E. granulosus DNA in 23.8% of serum samples from CE patients confirmed that E. granulosus releases cell-free DNA into the circulatory system, but quantities may be inadequate for the diagnosis of CE. Genotype G1 predominance suggests the sheep-dog cycle as the primary route of human infection.
Klíčová slova:
DNA – Sequence alignment – Polymerase chain reaction – Mitochondria – Multiple alignment calculation – Nucleotide sequencing – Echinococcosis – Cestodes
Zdroje
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