Developing Lateral Flow Assay Based Rapid Diagnosis Kit for Serologic Diagnosis of Cattle Brucellosis

Yıl 2018, Cilt: 29 Sayı: 1, 13 - 16, 30.04.2018

Osman Yaşar Tel Oktay Keskin , Sevil Erdenliğ Gürbilek

Öz

Brusellosis caused by microorganisms of Brucellagenus is a worldwide zoonotic infection seen in livestock and has serious risk for human and animal health and negative effect on animal production. The disease agents infect especially genital organs like uterus, testes, mammary glands and cause abortion and infertility in livestock. The aim of this study was to develop a rapid lateral flow test (LTF) for diagnosis cattle brucellosis. Sensitivity, specificity and diagnostic accuracy of LFT were evaluated by comparing the results with Rose Bengal Plate Test (RBPT), complement fixation test (CFT) and i-ELISA. Out of 91 serum samples 45 (49.5%), 44 (48.3%), 37 (40.7%) and 34 (37.4%) were found positive by RBPT, i-ELISA, CFT and LFT, respectively. Based on the results by CFT as reference test,  sensitivity and specificity of LFT were found 92% and  100%, respectively. As conclusion,  LFT and RBPT gave similar results and it was thought that LFT is an easy, rapid and specific screening test for diagnosis cattle brucellosis in the field.

Anahtar Kelimeler

Lateral Flow Assay, Cattle, Serologic diagnosis, Brucellosis

Sığır Brusellozunun Serolojik Teşhisinde Lateral Flow Temelli Hızlı Tanı Kiti Geliştirilmesi

Öz

Bruselloz, sığır, koyun, keçi, domuz ve koç gibi çiftlik hayvanlarında brusella grubu mikroorganizmaların sebep olduğu özellikle, testis, meme, uterus gibi genital organlara yerleşerek, yavru atmalara ve infertiliteye neden olan dünyanın birçok ülkesinde görülen insan ve hayvan sağlığını tehdit eden ve hayvansal üretim üzerine negatif etkileri olan önemli bir zoonozdur. Bu çalışma ile sığır brucellozisinin serolojik olarak çabuk tanısında kullanılacak bir LFT (Lateral flow test)’i geliştirmek amaçlandı. LFT’nin sensitivite, spesifisite ve tanısal doğruluğu, RBPT (Rose Bengal Plate Test), KFT (Komplement Fikzasyon Test) ve i-ELISA ile karşılaştırmalı olarak değerlendirildi. İncelen 91 serum örneğinin 44 (%48.3)’ü i-ELISA ile 34 (%37.4)’ü LFT, 45’i (%49.5) RBPT ile 37 (%40.7)’si KFT ile pozitif olarak saptandı. Komplement fikzasyon testi referans test kabul edilerek, LFT’nin sensitivitesi %92 ve spesifitesi %100 olarak bulundu. Sonuç olarak, LFT’nin, sığır brusellozun serolojik teşhisinde sahada kolayca uygulanabilecek hızlı sonuç alınan spesifik bir test olduğu sonucuna varıldı.

Anahtar Kelimeler

Lateral Flow Test, Sığır, Serolojik teşhiş, Bruselloz

Kaynakça

• Abdoel T, Dias IT, Cardoso R, Smits HL (2008). Simple and rapid field tests for brucellosis in livestock. Vet Microbiol, 30(3-4), 312-319. Alton GG (1990). Brucella melitensis. In: Animal Brucellosis, Nielsen K, Duncan JR (Eds), CRC Press, Boca Raton, Florida, USA, 383–409. Alton GG, Jones LM, Angus RD, Verger JM (1988). Techniques for the Brucellosis Laboratory. Inra, Paris; pp 63-136. Aydın E (2012). Brusella Enfeksiyonlarının Epidemiyolojisi. IV. Türkiye Zoonotik Hastalıklar Sempozyumu (Gıda Kaynaklı Zoonozlar), Sempozyum kitabı, 57-62. Blasco JM, Garin-Bastuji B, Marin CM, Gerbier G, Fanlo J, Jimenez De Bagues MP, Cau C (1994). Efficacy of different rose bengal and complement fixation antigens for the diagnosis of Brucella melitensis infection in sheep and goats. Vet Rec, 134, 415–420. Cheng S, Sun J, Yang J, Lv J, Wu F, Lin Y, Liao L, Ye Y, Cao C, Fang L, Hua Q (2017). A new immunoassay of serum antibodies against peste des petits ruminants virus using quantum dots and a lateral-flow test strip. Anal Bioanal Chem, 409(1), 133-141. Cox CR, Jensen KR, Mondesire RR, Voorhees KJ (2015). Rapid detection of Bacillus anthracis by γ phage amplification and lateral flow immunochromatography. J Microbiol Methods, 118, 51-56. Corbel MJ (1997). Brucellosis: an overview. Emerg Infect Dis, 3(2), 213-221. Frens G (1973). Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature, 241(105), 20-22. Greenfield RA, Drevets DA, Machado LJ, Voskuhl GW, Cornea P, Bronze MS (2002). Bacterial pathogens as biological weapons and agents of bioterrorism. Am J Med Sci, 323, 299-315. Irmak H, Buzgan T, Evirgen O, Akdeniz H, Demiroz AP, Abdoel TH, Smits HL (2004). Use of the Brucella IgM and IgG flow assays in the serodiagnosis of human brucellosis in an area endemic for brucellosis. Am J Trop Med Hyg, 70, 688-694. Kaewphinit T, Arunrut N, Kiatpathomchai W, Santiwatanakul S, Jaratsing P, Chansiri K (2013) Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples. Biomed Res Int. Vol: 2013, ID: 926230, 1-6. Kaltungo BY, Saidu SNA, Sackey AKB, Kazeem HM (2013). Serological evidence of brucellosis in goats in Kaduna North senatorial district of Kaduna State, Nigeria. ISRN Veterinary Science, Vol: 2013, ID: 963673, 1-6. McGiven J, Tucker J, Perrett L, Stack J, Brew S, MacMillan A (2003). Validation of FPA and cELISA for the detection of antibodies to Brucella abortus in cattle sera and comparison to SAT, CFT, and iELISA. J Immunol Methods, 278(1), 171-178. Nicoletti P (1969). Further evaluations of serologic test procedures used to diagnose brucellosis. Amer J Vet Res, 30(10), 1811-1816. Nielsen K (2002). Diagnosis of Brucellosis by serology. Vet Microbiol, 90(1), 447-459. OIE (2009). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Sixth Edition, Caprine And Ovine Brucellosis Page:1022-1030. Ruppanner R, Meyer M, Willeberg P, Behymer D (1980). Comparison of the enzyme-linked immunosorbent assay with other tests for Brucellosis, using sera from experimentally infected heifers. Am J Vet Res, 41(8), 1329-1332. Saha PK, Ali MH, Rahman MB, Islam MA (2010). Determination of sensitivity and specificity of in-house sandwich ELISA for the detection of infectious bursal disease viruses. Bangl J Vet Med, 8(2), 97-106. Saravi MA, Wright PF, Gregoret RJ, Gall DEJ (1995). Comparative performance of the enzyme-linked immunosorbent assay (ELISA) and conventional assays in the diagnosis of bovine brucellosis in Argentina. Vet immunol and Immunopathology, 47(1), 93-99. Shome R, Filia G, Padmashree BS, Krithiga N, Sahay S, Triveni K, Rahman H (2015). Evaluation of lateral flow assay as a field test for investigation of brucellosis outbreak in an organized buffalo farm: A pilot study. Veterinary world, 8(4), 492-496 Smits HL, Abdoel TH, Solera J, Clavijo E, Diaz R (2003). Immunochromatographic Brucella-specific immunoglobulin M and G lateral flow assays for rapid serodiagnosis of human brucellosis. Clin Diagn Lab Immunol, 10, 1141-1146. T.C. Sağlık Bakanlığı (2014). 2004-2006 İsatistikler / Temel Sağlık Hizmetleri Genel Müdürlüğü çalışma yıllığı. http://www.saglik.gov.tr/ TR/belge/1-2952/istatistik-yilliklari.html, Erişim tarihi: 18.03.2014. Wong R C, Tse HY (2009). Lateral Flow Immunoassay. Humana Press. New York, USA. Yüce A, Alp-Çavuş S (2006). Türkiye’de bruselloz: Genel bakış. Klin Derg, 19(3), 87-97.