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2017 | 20 | 2 |
Tytuł artykułu

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

Treść / Zawartość
Warianty tytułu
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.
Słowa kluczowe
Wydawca
-
Rocznik
Tom
20
Numer
2
Opis fizyczny
p.355-362,fig.,ref.
Twórcy
autor
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
Bibliografia
  • Andersen HJ, Pedersen LH, Aarestrup FM, Chriél M (2003) Evaluation of the surveillance program of Streptococcus agalactiae in Danish dairy herds. J Dairy Sci 86: 1233-1239.
  • Baseggio N, Mansell PD, Browning JW, Browning GF (1997) Strain differentiation of isolates of streptococci from bovine mastitis by pulsed-field gel electrophoresis. Mol Cell Probes 11: 349-354.
  • Boddie RL, Nickerson SC (1996) Efficacy of teat dips containing a hypochlorous acid germicide against experimental challenge with Staphylococcus aureus and Streptococcus agalactiae. J Dairy Sci 79: 1683-1688.
  • Boddie RL, Nickerson SC (2002) Reduction of mastitis caused by experimental challenge with Staphylococcus aureus and Streptococcus agalactiae by use of a quaternary ammonium and halogen-mixture teat dip. J Dairy Sci 85: 258-262.
  • Bosward KL, House JK, Deveridge A, Mathews K, Sheehy PA (2016) Development of a loop-mediated isothermal amplification assay for the detection of Streptococcus agalactiae in bovine milk. J Dairy Sci 99: 2142-2150
  • Bramley AJ, Schultze WD (1991) Effect of milking without pulsation on teat duct colonization with Streptococcus agalactiae and penetrability to endotoxin. J Dairy Sci 74: 2982-2988.
  • Dogan B, Schukken YH, Santisteban C, Boor KJ (2005) Distribution of serotypes and antimicrobial resistance genes among Streptococcus agalactiae isolates from bovine and human hosts. J Clin Microbiol 43: 5899-5906.
  • Duarte RS, Miranda OP, Bellei BC, Brito MA, Teixeira LM (2004) Phenotypic and molecular characteristics of Streptococcus agalactiae isolates recovered from milk of dairy cows in Brazil. J Clin Microbiol 42: 4214-4222.
  • Jzrgensen HJ, Nordstoga AB, Sviland S, Zadoks RN, Sølverød L, Kvitle B (2016) Streptococcus agalactiae in the environment of bovine dairy herds-rewriting the textbooks? Vet Microbiol 184: 64-72.
  • Keefe GP (1997) Streptococcus agalactiae mastitis: a review. Can Vet J 38: 429-437.
  • Mahmmod YS, Klaas IC, Katholm J, Lutton M, Zadoks RN (2015) Molecular epidemiology and strain-specific characteristics of Streptococcus agalactiae at the herd and cow level. J Dairy Sci 98: 6913-6924.
  • Meiri-Bendek I, Lipkin E, Friedmann A, Leitner G, Saran A, Friedman S (2002) A PCR-based method for the detection of Streptococcus agalactiae in milk. J Dairy Sci 85: 1717-1723.
  • Pinto TC, Costa NS, Corréa AB, de Oliveira IC, de Mattos MC, Rosado AS (2014) Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae. Braz J Microbiol 45: 785-789.
  • Pinto TC, Costa NS, Vianna Souza AR, Silva LG, Corréa AB, Fernandes FG (2013) Distribution of serotype and evaluation of antimicrobial susceptibility among human and bovine Streptococcus agalactiae strains isolated in Brazil between 1980 and 2006. Braz J Infect Dis 17: 131-136
  • Sukhnanand S, Dogan B, Ayodele MO, Zadoks RN, Craver MP, Dumas NB (2005) Molecular subtyping and characterization of bovine and human Streptococcus agalactiae isolates. J Clin Microbiol 43: 1177-1186.
  • Verhoeven PO, Noyel P, Bonneau J, Carricajo A, Fonsale N, Ros A (2014) Evaluation of the new brilliance GBS chromogenic medium for screening of Streptococcus agalactiae vaginal colonization in pregnant women. J Clin Microbiol 52: 991-993.
  • Vidanarachchi JK, Li S, Lundh DS, Johansson M (2015) Short communication: Lipolytic activity on milk fat by Staphylococcus aureus and Streptococcus agalactiae strains commonly isolated in Swedish dairy herds. J Dairy Sci 98: 8560-8564.
  • Vidová B, Chotár M, Godány A (2009) N-terminal anchor in surface immunogenic protein of Streptococcus agalactiae and its influence on immunity elicitation. Folia Microbiol (Praha) 54: 161-166.
  • Wang D, Liu Y (2015) Development of Primer Sets for Loop-Mediated Isothermal Amplification that Enables Rapid and Specific Detection of Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae. Int J Environ Res Public Health 12: 5735-5742.
  • Wang D, Wang Z, Yan Z, Wu J, Ali T, Li J (2015a) Bovine mastitis Staphylococcus aureus: Antibiotic susceptibility profile, resistance genes and molecular typing of methicillin-resistant and methicillin-sensitive strains in China. Infect Genet Evol 31C: 9-16.
  • Wang J, Chen J, Wei F, Dong Y, Zhu L, Han W (2015b) Prokaryotic Expression of Truncated S1 Protein of Porcine Epidemic Diarrhea Virus and Production of Monoclonal Antibodies to Recombinant Protein. Monoclon Antib Immunodiagn Immunother 5: 327-333.
  • Zanardi G, Caminiti A, Delle Donne G, Moroni P, Santi A, Galletti G (2014) Short communication: comparing real-time PCR and bacteriological cultures for Streptococcus agalactiae and Staphylococcus aureus in bulk-tank milk samples. J Dairy Sci 97: 5592-5598.
Typ dokumentu
Bibliografia
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Identyfikator YADDA
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