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Detection of Toxocara canis DNA in tissues of experimentally infected mice

Micaele, Quintana de Moura, Marcia, Raquel Pegoraro de Macedo, da Silva Terto, Wesley Douglas, da Costa Avila, Luciana Farias, Fabio, Pereira Leivas Leite, Carlos, Jaime Scaini, Natália, Berne Pinto, Gabriela, de Almeida Capella, Adriane, Leites Strothmann, Marcos, Marreiro Villela, Maria, Elisabeth Aires Berne
Acta tropica 2018
DNA, Toxocara canis, acute course, blood sampling, blood serum, brain, digestion, eggs, enzyme-linked immunosorbent assay, etiological agents, helminths, infectious diseases, internal transcribed spacers, larvae, liver, lungs, mice, polymerase chain reaction, signs and symptoms (animals and humans), tissues, toxocariasis
The main etiological agent of toxocariasis is the helminth Toxocara canis. Several difficulties are found in the diagnosis of this disease, because of nonspecific clinical signs and possible cross-reactions that may occur in the available test, the indirect ELISA. Therefore, molecular diagnosis has been indicated as an alternative to conventional diagnosis. The purpose of this study was to evaluate the polymerase chain reaction (PCR) technique for the identification of T. canis in tissues of experimentally infected mice. To this end, nine mice were inoculated with 1500 embryonated eggs and were divided into two groups, the first euthanized 48 h (G1) and the other 30 days post inoculation (G2). Lungs, brain, liver and blood were collected from all the animals for DNA Extraction and tissue digestion, also was collected blood samples for DNA extraction and ELISA test (serum). Toxocara canis DNA was identified in all the inoculated animals using the ITS-2 target gene. The PCR test successfully identified the parasite in the brain, lung and liver of the animals euthanized 48 h PI and 30 days PI. This technique yielded good results in the identification of the parasite in the brain, being more sensitive than the method for the recovery of larvae, in the group with acute infection (48 h PI). The infection was confirmed by PCR within 48 h after infection, while the ELISA indicated serological conversion occurred only 14 days after inoculation. This study demonstrates the ability of PCR to identify T. canis in the liver, lungs and brain during acute and chronic infection.