In vivo Study of Nano Chitosan Beads-Based ELISA versus Traditional Sandwich ELISA for the Early Diagnosis of Trichinosis.

Abstract

Human trichinosis is a serious foodborne parasitic zoonosis. Diagnosing human trichinosis is usually difficult due to the nonspecific clinical picture and the limited effectiveness of serological tests in acute infections. While ELISA can detect circulating Trichinella antigens, aiding in early diagnosis, its sensitivity may be low. The application of nanoparticles can improve the sensitivity of ELISA and allow a specific early diagnosis of the disease. This work compares the nano chitosan beads-based ELISA (NCSB-ELISA) and traditional sandwich ELISA for the detection of circulating Trichinella spiralis (T. spiralis) crude extract-antigen (CEA) in serum samples of experimentally infected mice. Fifty-seven mice included in this study were classified into 3 groups: T. spiralis infected group (Group I) (36 mice), which was equally subdivided into six subgroups according to the time of sacrifice (6, 8, 10, 12, 14 and 16) day post-infection (dpi), cross-reactivity group (Group II) (9 mice) and negative control group (Group III) (12 mice). T. spiralis AW-CEA prepared from the adult worms were used to produce anti- T. spiralis IgG-polyclonal antibodies in rabbits; these antibodies were utilized to detect AW-CEA in serum samples by traditional sandwich ELISA and NCSB-ELISA. Using NCSB-ELISA, T. spiralis AW-CEA was detected in sera collected at 8 dpi, with a sensitivity of 50% and a specificity of 100%. Meanwhile, traditional sandwich ELISA could not detect the antigen at the same time interval. Both ELISA techniques were able to detect the antigen in samples collected at 10, 12, 14 and 16 dpi with a sensitivity of 16.67%, 50%, 67.67% and 83.67%, respectively, for traditional sandwich-ELISA and a specificity of 100% at 10, 12 and 14 dpi while at 16 dpi specificity was decreased to 90.91%. In contrast, the sensitivity of NCSB-sandwich ELISA on the same days was 66.67%, 83.34%, 100% and 100%, respectively, with a specificity of 100% at all days. False positive detection of T. spiralis AW-CEA in the serum of mice in GII was recorded on day 16 pi by only traditional sandwich ELISA. This study concluded that NCSB-ELISA is a promising and sensitive technique for the early and specific diagnosis of acute trichinosis in an animal model.