ABSTRACT. Cysticercosis is a parasitic infection that causes severe economic and public health problems. The overall incidence of infection in sheep slaughtered in Cairo, Egypt during meat inspection was 31.22% and precisely 19.72% for Cysticercus tenuicollis and 11.50% for C. ovis. Sera collected from infected animals used to evaluate the diagnostic efficacy of the extracted antigens using ELISA. The sensitivity of the test was 100% for both C. tenuicollis and C. ovis, while the specificity was 80.26% and 87.03%, respectively. Using EITB the fractions of 36 KDa and 23 KDa appears to be specific for diagnosis of C. tenuicollis; while, 77 KDa and 73 KDa were specific for diagnosis of C. ovis. Moreover, several protein fractions which were detected in all antigens extracted didn't react with its target sera but at the same time did react specifically versus sera from animals infected with another cysticerci. Those fractions are considered as common immunogenic fractions between different cysticerci, so strictly identified specific fractions must to be used for diagnosis of Cysticerci infection to avoid cross reactions with other cysticerci antibodies.
SummaryToxocara canis of dogs and Toxocara vitulorum of cattle and buffalo are nematode parasites that cause serious economic and public health problems all over the world. This study aims to provide molecular data to identify and distinguish between Toxocara spp. from dogs, cattle and buffalo in Egypt. Moreover, constructing a phylogeny and phylogenetic relationships among these Toxocara spp. were performed through an analytic study of ATPase-6, a mitochondrial gene; 12S, small subunit ribosomal RNA gene and ITS-2, the second internal transcribed spacer nuclear ribosomal gene. T. vitulorum from cattle and buffalo were found to be almost identical. The ATPase- 6 and 12S regions showed 87.78 % and 90.38 % nucleotide similarity between T. canis and T. vitulorum, while for the ITS-2 region, only 78.38 % was found. Analysis of the three studied genes revealed that each Toxocara spp. has distinct molecular characteristics. Moreover, it was revealed that these genes, especially the ITS-2 gene, are useful and sensitive molecular markers for classifying and studying the phylogenetic analysis and relationships among closely related Toxocara spp. All sequences obtained in this study were registered in the GenBank under the accession numbers: MG214149 -MG214157.
Clinicians face significant problems in the diagnosis of zoonotic coenurosis. The current study aimed to develop an improved dot-Enzyme-linked-immunosorbent assay (dot-ELISA) for the diagnosis of zoonotic coenurosis using sheep Coenurus cerebralis scolices purified antigen (CcS-Ag) and to compare the obtained results with those of indirect ELISA and Enzyme-linked immunoelectrotransfer blot technique (EITB). Sera were collected from humans and sheep infected or suspected of infection with Coenurus cerebralis, control cases, and cases infected with other parasites. The CcS-Ag was proved to be the most specific antigen. This antigen was fractionated, and its specific polypeptides against anti-C. cerebralis antibodies (ACc-Ab) were identified using EITB. Fractions at the molecular weight (MW) of 48 and 58 kDa were proved as the only specific ones, eluted from the gel and concentrated, then dotted on the NC sheet as pooled antigen before its evaluation in the diagnosis of infection using dot-ELISA. Dot-ELISA demonstrated absolute 100% sensitivity and 100% specificity as recorded by EITB, compared to both fractions on a nitrocellulose (NC) sheet using surgically proved infected human or sheep sera as a gold standard. Diagnosis by ELISA using crude CcS-Ag revealed similar sensitivity but lower specificity (75%). The diagnostic accuracy of dot-ELISA was proved by comparing its results with postmortem data obtained post slaughtering of 20 suspected sheep and patients investigated by computed tomography (CT) and magnetic resonance imaging (MRI). In conclusion, the selection of specific fractions after EITB to be used in dot-ELISA improved the diagnostic value of the test as a diagnostic tool gathering the benefits of ELISA and EITB.
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