Foot and mouth disease (FMD) is highly infectious, contractable disease of ungulates and is associated with cattle which causes high economic losses in livestock industry. Virtual control of FMD requires specific and sensitive point-of-care diagnostic tools to eradicate the disease spreading. So far, the diagnostic tools used for Foot and Mouth Disease Virus (FMDV) are molecular based assays which are more expensive. The main objective of this research study is to develop and evaluate point-of-care test for rapid detection of FMDV in animals. For this study, a highly specific and sensitive FMD nonstructural protein (NSP) antibody rapid immunodiagnostic assay was developed using recombinant 3ABC (r3ABC) protein of FMDV for the detection of antibodies against FMDV and compared with commercial ELISA. The FMDV 3ABC gene was cloned into pET28a (+) vector and the gene product was expressed in E. coli BL21 cells. The expressed r3ABC protein was detected by SDS-PAGE analysis which resulted in a protein band with approximate molecular weight of 60 kDa. Purified r3ABC antigen was used as a detection reagent in rapid Lateral Flow assay (LFA). The diagnostic Assay was performed with 33 reference and 380 field samples and results showed that 94% sensitivity and 98.9% specificity. This study revealed that successful expression of 3ABC gene and diagnostic assay development lead to the identification of diseased animals. It further demonstrated that LFA as potential diagnostic tool for the point-of-care diagnosis of FMDV in large herds within limited time.
Salmonella enteritidis is a most important pathogenic bacterium of avian and mammals. Salmonella Enteritidis is the main cause of Salmonellosis in poultry flocks. S. enteritidis majorly infects the chicks, eggs and vertically transmitted to their off springs. The majority of the food infections to the humans are caused by salmonella by eating chicken meat and eggs. Monitoring of poultry farms with the bacteriological methods were time consuming and labour intensive process. The present study was development an in-house indirect enzyme linked immunesorbent assay (iELISA) for the detection of antibodies against Salmonella enteritidis in chicken serum samples. For detection of antibodies, Salmonella enteritidis LPS was used as antigen and rabbit anti chicken IgG HRP was used as the secondary antibody to detect antibodies against Salmonella enteritidis. The developed in-house ELISA was compared with the Rapid plate agglutination test. The purified LPS antigen 200ng/well, test sample serum at a dilution of 1:100 and rabbit anti chicken IgG HRP 1:10000 were used as optimal concentration of the assay and OD was measured at 450nm. A total of 1020 chicken serum samples were collected and performed the assay along with known Positive and negative controls. Out of these samples 592 and 566 samples were seropositive with iELISA and RPA respectively. Out of 1020 samples 58% samples shown positive immune response with iELISA and 55.6% samples were shown positive immune response to Rapid plate agglutination assay. The major prevalence of SE antibodies against SE antigen were shown in 20-25 weeks birds was 65.5%.The findings suggested that an in-house indirect ELISA based on S.enteritidis LPS can be a useful as a rapid and sensitive assay for the detection of antibodies to S.enteritidis and can be best assay for regular monitoring of Salmonella Enteritidis infection in flocks.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.