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Evaluation of Usage Indirect ELISA Using Antigens From Two Different Brucella Strains in Serological Diagnosis of B. canis Infection

Year 2016, Volume: 27 Issue: 3, 135 - 139, 20.12.2016

Abstract

The aim of this study was to develop two
indirect ELISAs (i-ELISA) using heat-extracted antigens prepared with the B. canis M- and B. abortus RB51 strains for the serologic diagnosis of B. canis infection in dogs. Reference
sera consisted of 16 agar gel immunodiffusion (AGID)-positive sera from animals
with clinical signs and positive bacterial isolation and 24 AGID-negative sera
from healthy animals with no history or evidence of canine brucellosis were
used to assess diagnostic sensitivity and specificity of the tests. Using
validation panel, the cut-off values of
OD450 0.243
and 0.354 were selected to give 100 % diagnostic specificity and sensitivity
for B. abortus RB51 i-ELISA and B. canis M- i-ELISA, respectively. Based
on these cut-off values, 113 serum samples taken from dog shelters in Şanlıurfa
and Bursa metropolitan municipalities tested by rapid slide agglutination test
(RSAT) and both i-ELISAs. Of the 47 dog sera tested from Şanlıurfa, 6 (12.8%)
were positive by RSAT, 3 (6.4%) were positive by B. abortus RB51-i-ELISA and two of the samples (4.2%) were found
positive by B. canis M- i-ELISA. Of
the 66 serum samples from Bursa, 11 (16.6%) were found positive by RSAT, 5
(7.6%) and 4 (6.1%) of which were confirmed by B. abortus RB51 i-ELISA and B.
canis
M- i-ELISA, respectively. In conclusion, both B. abortus RB51 i-ELISA and B.
canis
M- i-ELISA yielded the same diagnostic specificity and sensitivity
with minor differences and it was thought that they could be used successfully
as a confirmatory test together with RSAT as a screening test.

References

  • Alton GG, Jones LM, Angus RD, Verger JM (1988). Brucella canis. In: Techniques for the Brucellosis Laboratory, Institut National de la Recherche Agronomique, Puplication Paris, France, pp; 169-174. Badakhsh FF, Carmicheal LE, Douglas JA (1982). Improved rapid slide agglutination test for presumptive diagnosis of canine brucellosis. J Clin Microbiol, 15, 286-289. Barrouin-Melo SM, Poester FP, Riberio MB, Alcantara AC, Aguiar PHP, Nascimento IL, Schaer RE, Nascimento RM, Freire SM (2007). Diagnosis of canine brucellosis by ELISA using an antigen obtained from wild Brucella canis. Res Vet Sci, 83, 340-346. Carmichael LE, Joubert JC (1987). A rapid slide agglutination test for the serodiagnosis of Brucella canis infection that employs a variant (M-) organism as antigen, Cornell Vet, 77, 3-12. Carmichael LE, Kenney RM (1970). Canine brucellosis: the clinical disease, pathogenesis, and immune response. J Am Vet Med Assoc, 156, 1726. Carmichael LE (1966). Abortions in 200 beagles. J Am Vet Med Assoc, 149, 1126. Carmichael LE (1990). Brucella canis. In : Nielsen K, Duncan JR, eds. Animal Brucellosis, Boca Raton FL, CRC Press, pp; 335-350. Currier RW, Raithel WF, Martin RJ, Potter ME (1982). Canine brucellosis, J Am Vet Med Ass, 180, 132-133. Diker KS, İstanbulluoğlu E, Ayhan H, Soysal G (1984). A serosurvey of Brucella canis infections in man at Bursa district. Mikrobiyol Bult, 18, 203-207. Diker, KS, Aydın N, Erdeger J, Ozyurt M (1987). A serologic survey of dogs for Brucella canis and Brucella abortus and evaluation of mercaptoethanol microagglutination test. Ankara Univ Vet Fak Derg, 34, 268-277. Escobar GI, Boeri EJ, Ayala SM, Lucero NE (2010). The feasibility of using prepared with rough Brucella strains for diagnosis of canine brucellosis. Rev Argent Microbiol, 42, 35-40. Hollett RB (2006). Canine brucellosis: Outbreaks and compliance. Theriogenology, 66, 575–587. İstanbulluoğlu E, Diker, KS (1983). An serological analysis of Brucella canis. Ankara Univ Vet Fak Derg, 30, 14-18. Lopez G, Ayala SM, Efron AM, Gomez CF, Lucero NE (2009). A serological and bacteriological survey of dogs to detect Brucella infection in Lomas de Zamora Buenos Aires province. Rev Argent Microbiol, 41, 97-101. Lopez G, Ayala SM, Escobar GI, Lucero NE (2005). Use of Brucella canis antigen for detection of ovine serum antibodies against Brucella ovis. Vet Microbiol, 2005, 105, 181-187. Lucero NE, Escobar GI, Ayala SA, Lopez G (2002). Sensitivity and specificity of an indirect enzyme-linked immunoassay for the diagnosis of Brucella canis infection in dogs. J Med Microbiol, 51, 656-660. Mateu-de-antonio EM, Martin M, Casal J (1994). Comparison of serological tests used in canine brucellosis diagnosis. J Vet Diagn Invest, 6, 257-259. Mateu-de-antonio EM, Martin M, Soler M (1993). Use of indirect enzyme-linked immunosorbent assay with hot saline solution extracts of a variant (M-) strain of Brucella canis. Am J Vet Res, 54, 1043-1046. Moreno E, Jones LM, Berman DT (1984). Immunochemical characterization of Rough Brucella Lipopolysaccharides, Infect Immun, 43, 779-782. Nielsen K, Smith P, Conde S, Dragide Benitez G, Gall D, Halbert G, Kenny K, Massengill C, Muenks Q, Rojas X, Perez B, Samartino L, Silva P, Tollersrud T, Lolley M (2004). Rough lipopolysaccharide of Brucella abortus RB51 as a common antigen for serological detection of B. ovis, B. canis, and B. abortus RB51 exposure using indirect enzyme immunoassay and fluorescence Polarization assay. J Immunoass Immunoch, 25, 171-182. Nielsen K, Smith P, Yu WL, Rojas X, Perez B, Conde S, Samartino L, Robles C (2007). Detection of ovine antibody to Brucella ovis by indirect enzyme immunoassay. J Immunoass Immunoch, 28, 243-250. Oliveira MZD, Vale V, Keid L, Freire SM, Meyer R, Portela R, Barrouin-Melo SM (2011). Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis. Res Vet Sci, 90, 425-431. Öncel T, Akan M, Sareyyüpoğlu B, Tel OY, Çiftçi A (2005). Seroprevalance of Brucella canis infection of dogs in two provinces in Turkey. Turk J Vet Anim Sci, 29, 779-783. Pickerill PA (1970). Comments on epizootiology and control of canine brucellosis. J Am Vet Med Assoc, 156, 1741-1742. Schoonjans F, Zalata A, Depuyd, CE, Comhaire FH (1995). MedCalc:a new computer program for medical statistics. Comput. Methods Programs Biomed, 48, 257-262. Uçan US, Aras Z, Semacan A (2010). Serodiagnosis of Brucella canis infection in dogs by a dipstick enzyme immunoassay. Eurasian J Vet Sci, 26, 109-112. Wanke MM, Delpino V, Baldi PC (2002). Comparative performance of tests using cytosolic or outer membrane antigens of Brucella for the serodiagnosis of canine brucellosis. Vet Microbiol, 88, 367-375. Wanke, MM (2004). Canine brucellosis. Anim Reprod Sci, 82-83, 195–207. Yardımcı H, Akan M, Atasever A (1998). Brucella canis’in biyolojik materyallerdeki yaşam süresi. YYÜ Vet Fak Derg, 9, 77-79. Zoha SJ, Carmicheal LE (1981). Properties of Brucella canis surface antigens associated with colonial mucoidness. Cornell Vet, 71, 428-438. Zoha SJ, Carmicheal LE (1982). Properties of cell wall antigens of virulent Brucella canis and a less mucoid variant of reduced pathogenicity. Am J Vet Res, 43, 171-174.

Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması

Year 2016, Volume: 27 Issue: 3, 135 - 139, 20.12.2016

Abstract

Bu
çalışmada, B. canis infeksiyonunun
serolojik teşhisi için B. canis M- ve
B.abortus RB51 suşlarından hazırlanan
antijenlerin kullanıldığı indirekt ELISA (i-ELISA) geliştirilmesi
amaçlanmıştır. B. canis infeksiyonu
etken izolasyonu ve agar jel immunodiffuzyon (AGID) testi ile doğrulanmış 16
adet pozitif serum ve B. canis
infeksiyonu yönünden hiçbir klinik belirti göstermeyen AGID negatif 24 adet
serum testin duyarlılığı ve özgüllüğünü saptamak için referans serumlar olarak
kullanılmıştır. Validasyon paneli kullanılarak, tanısal duyarlılık ve özgüllüğü
%100 verecek eşik değeri B. abortus
RB51 i-ELISA için OD450 0.243 ve B.
canis
M- i-ELISA için OD450 0.354 olarak belirlenmiştir.
Saptanan eşik değer göz önüne alınarak, Şanlıurfa ve Bursa Büyükşehir
Belediyesi Köpek Barınaklarından temin edilen 113 adet köpek serumu çabuk lam
aglütinasyon testi (ÇLAT) ve geliştirilen iki i-ELISA prototipi ile test
edilmiştir.  Şanlıurfa ilinden temin
edilen 47 köpek serumundan 6’sı (%12.8) ÇLAT ile, 3’ü (%6.4) B. abortus RB51 i-ELISA ile ve ikisi
(%4.2) de B. canis M- i-ELISA ile
pozitif bulunmuştur. Bursa ilinden temin edilen 66 adet köpek serumundan 11’i
(%16.6) ÇLAT ile, 5’i (%7.6) B. abortus
RB51 i-ELISA ile ve 4’ü (%6.1) de B.
canis
M- i-ELISA ile pozitif bulunmuştur. Sonuçta B. abortus RB51 i-ELISA ve B.
canis
M- i-ELISA’nın küçük farklılıklar ile benzer duyarlılık ve özgüllük
gösterdikleri ve bu testlerin köpeklerde B.
canis
infeksiyonunda doğrulayıcı test olarak ÇLAT ile birlikte başarılı bir
şekilde kullanılabileceği kanısına varılmıştır.

References

  • Alton GG, Jones LM, Angus RD, Verger JM (1988). Brucella canis. In: Techniques for the Brucellosis Laboratory, Institut National de la Recherche Agronomique, Puplication Paris, France, pp; 169-174. Badakhsh FF, Carmicheal LE, Douglas JA (1982). Improved rapid slide agglutination test for presumptive diagnosis of canine brucellosis. J Clin Microbiol, 15, 286-289. Barrouin-Melo SM, Poester FP, Riberio MB, Alcantara AC, Aguiar PHP, Nascimento IL, Schaer RE, Nascimento RM, Freire SM (2007). Diagnosis of canine brucellosis by ELISA using an antigen obtained from wild Brucella canis. Res Vet Sci, 83, 340-346. Carmichael LE, Joubert JC (1987). A rapid slide agglutination test for the serodiagnosis of Brucella canis infection that employs a variant (M-) organism as antigen, Cornell Vet, 77, 3-12. Carmichael LE, Kenney RM (1970). Canine brucellosis: the clinical disease, pathogenesis, and immune response. J Am Vet Med Assoc, 156, 1726. Carmichael LE (1966). Abortions in 200 beagles. J Am Vet Med Assoc, 149, 1126. Carmichael LE (1990). Brucella canis. In : Nielsen K, Duncan JR, eds. Animal Brucellosis, Boca Raton FL, CRC Press, pp; 335-350. Currier RW, Raithel WF, Martin RJ, Potter ME (1982). Canine brucellosis, J Am Vet Med Ass, 180, 132-133. Diker KS, İstanbulluoğlu E, Ayhan H, Soysal G (1984). A serosurvey of Brucella canis infections in man at Bursa district. Mikrobiyol Bult, 18, 203-207. Diker, KS, Aydın N, Erdeger J, Ozyurt M (1987). A serologic survey of dogs for Brucella canis and Brucella abortus and evaluation of mercaptoethanol microagglutination test. Ankara Univ Vet Fak Derg, 34, 268-277. Escobar GI, Boeri EJ, Ayala SM, Lucero NE (2010). The feasibility of using prepared with rough Brucella strains for diagnosis of canine brucellosis. Rev Argent Microbiol, 42, 35-40. Hollett RB (2006). Canine brucellosis: Outbreaks and compliance. Theriogenology, 66, 575–587. İstanbulluoğlu E, Diker, KS (1983). An serological analysis of Brucella canis. Ankara Univ Vet Fak Derg, 30, 14-18. Lopez G, Ayala SM, Efron AM, Gomez CF, Lucero NE (2009). A serological and bacteriological survey of dogs to detect Brucella infection in Lomas de Zamora Buenos Aires province. Rev Argent Microbiol, 41, 97-101. Lopez G, Ayala SM, Escobar GI, Lucero NE (2005). Use of Brucella canis antigen for detection of ovine serum antibodies against Brucella ovis. Vet Microbiol, 2005, 105, 181-187. Lucero NE, Escobar GI, Ayala SA, Lopez G (2002). Sensitivity and specificity of an indirect enzyme-linked immunoassay for the diagnosis of Brucella canis infection in dogs. J Med Microbiol, 51, 656-660. Mateu-de-antonio EM, Martin M, Casal J (1994). Comparison of serological tests used in canine brucellosis diagnosis. J Vet Diagn Invest, 6, 257-259. Mateu-de-antonio EM, Martin M, Soler M (1993). Use of indirect enzyme-linked immunosorbent assay with hot saline solution extracts of a variant (M-) strain of Brucella canis. Am J Vet Res, 54, 1043-1046. Moreno E, Jones LM, Berman DT (1984). Immunochemical characterization of Rough Brucella Lipopolysaccharides, Infect Immun, 43, 779-782. Nielsen K, Smith P, Conde S, Dragide Benitez G, Gall D, Halbert G, Kenny K, Massengill C, Muenks Q, Rojas X, Perez B, Samartino L, Silva P, Tollersrud T, Lolley M (2004). Rough lipopolysaccharide of Brucella abortus RB51 as a common antigen for serological detection of B. ovis, B. canis, and B. abortus RB51 exposure using indirect enzyme immunoassay and fluorescence Polarization assay. J Immunoass Immunoch, 25, 171-182. Nielsen K, Smith P, Yu WL, Rojas X, Perez B, Conde S, Samartino L, Robles C (2007). Detection of ovine antibody to Brucella ovis by indirect enzyme immunoassay. J Immunoass Immunoch, 28, 243-250. Oliveira MZD, Vale V, Keid L, Freire SM, Meyer R, Portela R, Barrouin-Melo SM (2011). Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis. Res Vet Sci, 90, 425-431. Öncel T, Akan M, Sareyyüpoğlu B, Tel OY, Çiftçi A (2005). Seroprevalance of Brucella canis infection of dogs in two provinces in Turkey. Turk J Vet Anim Sci, 29, 779-783. Pickerill PA (1970). Comments on epizootiology and control of canine brucellosis. J Am Vet Med Assoc, 156, 1741-1742. Schoonjans F, Zalata A, Depuyd, CE, Comhaire FH (1995). MedCalc:a new computer program for medical statistics. Comput. Methods Programs Biomed, 48, 257-262. Uçan US, Aras Z, Semacan A (2010). Serodiagnosis of Brucella canis infection in dogs by a dipstick enzyme immunoassay. Eurasian J Vet Sci, 26, 109-112. Wanke MM, Delpino V, Baldi PC (2002). Comparative performance of tests using cytosolic or outer membrane antigens of Brucella for the serodiagnosis of canine brucellosis. Vet Microbiol, 88, 367-375. Wanke, MM (2004). Canine brucellosis. Anim Reprod Sci, 82-83, 195–207. Yardımcı H, Akan M, Atasever A (1998). Brucella canis’in biyolojik materyallerdeki yaşam süresi. YYÜ Vet Fak Derg, 9, 77-79. Zoha SJ, Carmicheal LE (1981). Properties of Brucella canis surface antigens associated with colonial mucoidness. Cornell Vet, 71, 428-438. Zoha SJ, Carmicheal LE (1982). Properties of cell wall antigens of virulent Brucella canis and a less mucoid variant of reduced pathogenicity. Am J Vet Res, 43, 171-174.
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Details

Primary Language Turkish
Journal Section Articles
Authors

Sevil Erdenliğ Gürbilek This is me

Oktay Keskin

Esra B Üyükçangaz This is me

Osman Yaşar Tel

Publication Date December 20, 2016
Submission Date March 31, 2016
Acceptance Date June 2, 2016
Published in Issue Year 2016 Volume: 27 Issue: 3

Cite

APA Erdenliğ Gürbilek, S., Keskin, O., Üyükçangaz, E. B., Tel, O. Y. (2016). Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması. Van Veterinary Journal, 27(3), 135-139.
AMA Erdenliğ Gürbilek S, Keskin O, Üyükçangaz EB, Tel OY. Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması. Van Vet J. December 2016;27(3):135-139.
Chicago Erdenliğ Gürbilek, Sevil, Oktay Keskin, Esra B Üyükçangaz, and Osman Yaşar Tel. “Brucella Canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması”. Van Veterinary Journal 27, no. 3 (December 2016): 135-39.
EndNote Erdenliğ Gürbilek S, Keskin O, Üyükçangaz EB, Tel OY (December 1, 2016) Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması. Van Veterinary Journal 27 3 135–139.
IEEE S. Erdenliğ Gürbilek, O. Keskin, E. B. Üyükçangaz, and O. Y. Tel, “Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması”, Van Vet J, vol. 27, no. 3, pp. 135–139, 2016.
ISNAD Erdenliğ Gürbilek, Sevil et al. “Brucella Canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması”. Van Veterinary Journal 27/3 (December 2016), 135-139.
JAMA Erdenliğ Gürbilek S, Keskin O, Üyükçangaz EB, Tel OY. Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması. Van Vet J. 2016;27:135–139.
MLA Erdenliğ Gürbilek, Sevil et al. “Brucella Canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması”. Van Veterinary Journal, vol. 27, no. 3, 2016, pp. 135-9.
Vancouver Erdenliğ Gürbilek S, Keskin O, Üyükçangaz EB, Tel OY. Brucella canis İnfeksiyonunun Serolojik Tanısında İki Farklı Brucella Suşundan Hazırlanan Antijenlerin İndirekt ELISA’da Kullanılabilirliğinin Araştırılması. Van Vet J. 2016;27(3):135-9.

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