Résumé
Background: Dirofilaria immitis is a cosmopolitan zoonotic, vector-borne parasite of carnivorous animals causing dirofilariasis in human beings. Common commercial serodiagnostic tests for canine dirofilariasis usually lead to different results in their sensitivity and specificity. The present study reports development of recombinant DgK [rDgK] antigen of D. immitis for accurate immunodiagnosis of D. Immitis-infected dogs using indirect ELISA test
Methods: The rDgK coding sequence was successfully sequenced, codon optimized and cloned in pET-24a[+] expression vector and then expressed in Escherichia coli. The recombinant DgK was affinity purified using Ni[+2] - charged HiTrap chelating column, followed by testing in Western blotting and enzyme-linked immunosorbent assays [ELISA] with dog sera from a dirofilariasis endemic area. The performance of rDgK ELISA was evaluated using 60 sera collected from suspected dogs, while molecular technique was used as a reference test
Results: Sera from positive control D. immitis infection produced a strong IgG antibody response to rDgK both in ELISA and Western blotting tests. The sensitivity and specificity related to diagnostic potential of rDgK for ELISA were 92.5% and 87.5%, respectively. The results of rDgK ELISA showed a high agreement [0.764] with molecular identification
Conclusions: The findings revealed that the developed new rDgK antigen is sensitive and specific for immunodiagnosis of canine dirofilariasis using ELISA test
Résumé
Aim: Detection of protein expression changes in human cystic echinococcosis sera by 2D gel electrophoresis
Background: Diagnosis and successful treatment of cystic echinococcosis [CE] is a major challenge, up to now. Identification of related expressed proteins using proteomics tools and bioinformatics analysis of patientsf sera have not been investigated, so far
methods: Sera from eight confirmed CE patients and three healthy controls were collected, tested by 2-DE for total protein separation of serum and analyzed using proteomics and bioinformatics methods. The gels were stained by Coomassie blue followed by scan imaging of the gels. The protein spots in each gel were analyzed using progenesis same spots software. Proteins names were obtained from TagIdent server
Results: A total of 263 protein spots with different expression were detected in both normal and diseased samples. Comparison between diseased and normal gels showed the expression of 45 up-regulated protein spots with fold.2 in diseased gel of which 10 were new proteins with statistical difference by normal gel [p-value<0.05]. On the other hand, the expression of 50 down-regulated protein spots were observed of which 11 proteins have been suppressed. Clustering of all detected sera proteins [263] using correlation analysis, divided the proteins into 2 clusters based on up-regulated and down-regulated expression of proteins. Clustering results were approved by principal component analysis [PCA]
Conclusion: Significant protein expression changes in human CE sera which is demonstrable by application of proteomics and bioinformatics analysis makes it an impressing tool for diagnosis of CE patients