Dataset Information

Optimization of In-House Indirect-ELISA & EITB Assays Employing Cysticercus cellulosae Antigens for Serological Detection and PCR Assays for Molecular Detection of Porcine Cysticercosis.

ABSTRACT:

Background

Porcine cysticercosis, caused by metacestodes of Taenia solium is an important neglected zoonosis. We evaluated the presence of anti-cysticercal antibodies and T. solium specific DNA in pig sera and blood samples respectively collected from Maharashtra, India.

Methods

A total of three antigens (Scolex Antigen (SA), Membrane Body Antigen (MBA) and Excretory-Secretory Antigen (ESA)) were prepared from metacestodes of T. solium and employed in an in-house developed indirect-IgG ELISA for serological screening of 1000 porcine sera samples at Department of Veterinary Public Health, Nagpur Veterinary College, Maharashtra, India. The ELISA positive sera samples were subjected to EITB Assay for detection of immunodominant peptides. An effort has been made for molecular detection of porcine cysticercosis by PCR assay targeting large subunit rRNA gene of T. solium from blood samples of the corresponding ELISA-positive pigs.

Results

The overall seroprevalence of porcine cysticercosis employing SA, MBA and ESA was 12.6%, 8.7% and 12.5% respectively. The lower and medium molecular weight peptides were the most frequently recognised in EITB assay. The numbers of bands recognised in EITB assay were observed to be proportionate with the corresponding ELISA O.D. values. An amplification product of 286 bp was observed in 22.98% (20/87), 30.35% (30/99) and 17.14% (12/70) of the sero-positive samples against SA, ESA and MBA respectively.

Conclusion

EITB still remains the gold standard serodiagnosis test for cysticercosis. The inclusion of a greater number of positive samples and purification of antigens may improve the diagnostic efficacy of the tests.

SUBMITTER: Satyaprakash K

PROVIDER: S-EPMC10183449 | biostudies-literature | 2023 Jan-Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Optimization of In-House Indirect-ELISA & EITB Assays Employing <i>Cysticercus cellulosae</i> Antigens for Serological Detection and PCR Assays for Molecular Detection of Porcine Cysticercosis.

Satyaprakash Kaushik K Khan Wiqar Ahmed WA Chaudhari Sandeep Prabhakarrao SP Shinde Shilpshri Vasant SV Kolte Sunil Waman SW Pansare Nilesh Ramchandra NR Likhite Amrut Vijay AV

Iranian journal of parasitology 20230101 1

<h4>Background</h4>Porcine cysticercosis, caused by metacestodes of <i>Taenia solium</i> is an important neglected zoonosis. We evaluated the presence of anti-cysticercal antibodies and <i>T. solium</i> specific DNA in pig sera and blood samples respectively collected from Maharashtra, India.<h4>Methods</h4>A total of three antigens (Scolex Antigen (SA), Membrane Body Antigen (MBA) and Excretory-Secretory Antigen (ESA)) were prepared from metacestodes of <i>T. solium</i> and employed in an in-ho ...[more]

PMID: 37197077

Similar Datasets

Project description:BackgroundA novel porcine deltacoronavirus (PDCoV), also known as porcine coronavirus HKU15, was reported in China in 2012 and identified in the U.S. in early 2014. Since then, PDCoV has been identified in a number of U.S. states and linked with clinical disease including acute diarrhea and vomiting in the absence of other identifiable pathogens. Since PDCoV was just recently linked with clinical disease, few specific antibody-based reagents were available to assist in diagnosis of PDCoV and limited serological capabilities were available to detect an antibody response to this virus. Therefore, the overall objective of this project was to develop and validate selected diagnostic reagents and assays for PDCoV antigen and antibody detection.ResultsThe nucleoprotein of PDCoV was expressed as a recombinant protein and purified for use as an antigen to immunize mice for polyclonal, hyperimmune sera and monoclonal antibody (mAb) production. The resulting mAbs were evaluated for use in fluorescent antibody staining methods to detect PDCoV infected cells following virus isolation attempts and for immunohistochemistry staining of intestinal tissues of infected pigs. The same antigen was used to develop serological tests to detect the antibody response to PDCoV in pigs following infection. Serum samples from swine herds with recent documentation of PDCoV infection and samples from expected naïve herds were used for initial assay optimization. The tests were optimized in a checkerboard fashion to reduce signal to noise ratios using samples of known status. Statistical analysis was performed to establish assay cutoff values and assess diagnostic sensitivities and specificities. At least 629 known negative serum samples and 311 known positive samples were evaluated for each assay. The enzyme linked immunosorbent assay (ELISA) showed diagnostic sensitivity (DSe) of 96.1% and diagnostic specificity (DSp) of 96.2%. The fluorescent microsphere immunoassay (FMIA) showed a DSe of 95.8% and DSp of 98.1%. Both ELISA and FMIA detected seroconversion of challenged pigs between 8-14 days post-infection (DPI). An indirect fluorescent antibody (IFA) test was also developed using cell culture adapted PDCoV for comparative purposes.ConclusionThese new, specific reagents and serological assays will allow for improved diagnosis of PDCoV. Since many aspects of PDCoV infection and transmission are still not fully understood, the reagents and assays developed in this project should provide valuable tools to help understand this disease and to aid in the control and surveillance of porcine deltacoronavirus outbreaks.