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2015 - 20162
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Author: RATNA KISHORE, G ×
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    ThesisItemOpen Access
    EVALUATION OF FLOW THROUGH ASSAY (FTA) AND INDIRECT ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR DETECTION OF OVINE FASCIOLOSIS
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P) INDIA, 2015-12) RATNA KISHORE, G; MALAKONDAIAH, P(MAJOR); SREE DEVI, C; SUBRAMANYAM, K V
    ABSTRACT: A study was under taken to develop immunodiagnostics for the detection ofantibodies to ovine fasciolosis. Two diagnostic antigens viz. excretory and secretoryantigen (ESA) and somatic antigen (SOA) were prepared from fasciolosis positive sheep and were evaluated by flow through assay (FTA) and enzyme linked immunosorbentassay (ELISA). The FTA was developed and compared with that of meat inspection and faecal examination results. The ELISA was standardized as confirmatory test for antibody detection in ovine fasciolosis and used to compare with FTA. A total of 350 sheep faecal samples collected from five districts of Andhra Pradesh were examined by standard coprological sedimentation method. Out of 350 samples, 20 samples were found positive for eggs of F. gigantica indicating 5.71 per cent of overall prevalence. Among five districts the rate of prevalence was high in Krishna district (10.0%) followed by Guntur (6.25%), East Godavari (2.0%), West Godavari (2.0%) and Chittoor (2.0%) districts. The highest prevalence of fasciolosis was recorded in adult sheep (6.57%) followed by young (5.17%) and lambs (2.78%). The prevalence was significantly higher in females (7.84%) as compared to males (2.74%). Both ESA and SOA at 550 ng/l and 150 ng/l concentrations gave optimum dot intensity in FTA, respectively. A total of 200 sheep serum samples were collected from five districts of Andhra Pradesh for evaluation of FTA and ELISA. Among the 200 serum samples tested, 74 (37.0%) and 38 (19.0%) samples gave positive reaction in FTA with ESA and SOA, respectively. In comparison with meat inspection and faecal examination, the ESA-FTA showed 100 per cent specificity, sensitivity and accuracy, with a K value of 0.95 indicating perfect agreement. Specificity, sensitivity and accuracy for SOA-FTA were 100, 90.9 and 95.2 per cent respectively, with a K value of 0.96 indicating perfect agreement. Out of 200 serum samples screened by ELISA, 118 (59.0%) and 130 (65.0%) gave positive reaction with ESA and SOA, respectively. The results with ESA-ELISA revealed 100 per cent specificity, sensitivity, and accuracy with K value of 0.95 indicating perfect agreement. The specificity, sensitivity and accuracy values with SOA-ELISA were 87, 100 and 93 per cent respectively. The FTA results were comparable with faecal examination and ELISA and could be easily performed even at room temperature within 3 min. that can be used for screening of serum samples at field level using ESA.
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    ThesisItemOpen Access
    EVALUATION OF FLOW THROUGH ASSAY (FTA) AND INDIRECT ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR DETECTION OF OVINE FASCIOLOSIS
    (SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502 (A.P) INDIA., 2016-12) RATNA KISHORE, G; MALAKONDAIAH, P (Major); SREE DEVI, C; SUBRAMANYAM, K.V
    ABSTRACT : A study was under taken to develop immunodiagnostics for the detection of antibodies to ovine fasciolosis. Two diagnostic antigens viz. excretory and secretory antigen (ESA) and somatic antigen (SOA) were prepared from fasciolosis positive sheep and were evaluated by flow through assay (FTA) and enzyme linked immunosorbent assay (ELISA). The FTA was developed and compared with that of meat inspection and faecal examination results. The ELISA was standardized as confirmatory test for antibody detection in ovine fasciolosis and used to compare with FTA. A total of 350 sheep faecal samples collected from five districts of Andhra Pradesh were examined by standard coprological sedimentation method. Out of 350 samples, 20 samples were found positive for eggs of F. gigantica indicating 5.71 per cent of overall prevalence. Among five districts the rate of prevalence was high in Krishna district (10.0%) followed by Guntur (6.25%), East Godavari (2.0%), West Godavari (2.0%) and Chittoor (2.0%) districts. The highest prevalence of fasciolosis was recorded in adult sheep (6.57%) followed by young (5.17%) and lambs (2.78%). The prevalence was significantly higher in females (7.84%) as compared to males (2.74%). Both ESA and SOA at 550 ng/l and 150 ng/l concentrations gave optimum dot intensity in FTA, respectively. A total of 200 sheep serum samples were collected from five districts of Andhra Pradesh for evaluation of FTA and ELISA. Among the 200 serum samples tested, 74 (37.0%) and 38 (19.0%) samples gave positive reaction in FTA with ESA and SOA, respectively. In comparison with meat inspection and faecal examination, the ESA-FTA showed 100 per cent specificity, sensitivity and accuracy, with a K value of 0.95 indicating perfect agreement. Specificity, sensitivity and accuracy for SOA-FTA were 100, 90.9 and 95.2 per cent respectively, with a K value of 0.96 indicating perfect agreement. Out of 200 serum samples screened by ELISA, 118 (59.0%) and 130 (65.0%) gave positive reaction with ESA and SOA, respectively. The results with ESA-ELISA revealed 100 per cent specificity, sensitivity, and accuracy with K value of 0.95 indicating perfect agreement. The specificity, sensitivity and accuracy values with SOA-ELISA were 87, 100 and 93 per cent respectively. The FTA results were comparable with faecal examination and ELISA and could be easily performed even at room temperature within 3 min. that can be used for screening of serum samples at field level using ESA.