Indirect enzyme-linked immunosorbent assay method based on Streptococcus agalactiae rSip-Pgk-FbsA fusion protein for detection of bovine mastitis

• Autorzy: Bu R.-E. , Wang I.-L. , Wu J.-H. , Xilin G.W. , Chen J.-L. , Wang H.
• Języki publikacji: EN

Abstrakty

EN

Objective: The aim of this study was to establish a rapid and accurate method for the detection of the Streptococcus agalactiae antibody (SA-Ab) to determine the presence of the bovine mastitis (BM)-causative pathogen. Methods: The multi-subunit fusion protein rSip-Pgk-FbsA was prokaryotically expressed and purified. The triple activities of the membrane surface-associated proteins Sip, phosphoglycerate kinase (Pgk), and fibronectin (FbsA) were used as the diagnostic antigens to establish an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of SA-Ab in BM. Results: The optimal antigen coating concentration was 2 μg/mL, the optimal serum dilution was 1:160, and the optimal dilution of the enzyme-labeled secondary antibody was 1:6000. The sensitivity, specificity, and repeatability tests showed that the method established in this study had no cross-reaction with antibodies to Streptococcus pyogenes, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis in the sera. The results of the sensitivity test showed that a positive result could be obtained even if the serum dilution reached 1:12,800, indicating the high sensitivity and good repeatability of the method. The positive coincidence rate of this method was 98.6%, which is higher than that of previous tests established with the Sip or Pgk mono-antigen fusion protein, respectively, demonstrating the relatively higher sensitivity of this newly established method. The detection rate for 389 clinical samples was 46.53%. Conclusions: The indirect ELISA method established in this study could provide a more accurate and reliable serological method for the rapid detection of S. agalactiae in cases of BM.

• Wydawca: -
• Czasopismo: Polish Journal of Veterinary Sciences
• Rocznik: 2017
• Tom: 20
• Numer: 2
• Opis fizyczny: p.355-362,fig.,ref.

Twórcy

autor

Bu R.-E.

• College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043, China
• Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention and Control the Pathogenic Bacteria in Milk, Tongliao 028043, China

autor

Wang I.-L.

• Key Lab of Prevention Veterinary and Animal Biological Technology, Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, Shandong Province, China
• Division of Animal Disease Detection, Shandong LVDU Bio-tech Co., Ltd. Binzhou 256600, Shandong Province, China

autor

Wu J.-H.

• College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043, China
• Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention and Control the Pathogenic Bacteria in Milk, Tongliao 028043, China

autor

Xilin G.W.

• College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043, China
• Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention and Control the Pathogenic Bacteria in Milk, Tongliao 028043, China

autor

Chen J.-L.

• Key Lab of Prevention Veterinary and Animal Biological Technology, Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, Shandong Province, China
• Division of Animal Disease Detection, Shandong LVDU Bio-tech Co., Ltd. Binzhou 256600, Shandong Province, China

autor

Wang H.

• College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043, China
• Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention and Control the Pathogenic Bacteria in Milk, Tongliao 028043, China

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Identyfikatory

DOI

10.1515/pjvs-2017-0043