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Abstract

Infectious Bovine Rhinotracheitis (IBR) is caused by Bovine Herpesvirus-1 (BHV-1) that shows a worldwide distribution and results in severe economic losses in the cattle industry. The aims of this study were to; detect the antibodies against the BHV-1 in cattle blood sera samples by the Enzyme-Linked Immunosorbent Assay (ELISA) and micro Neutralisation Test (mNT) and determine the specifity and sensitivity values of these serologic diagnostic tests. Blood samples were randomly collected from 301 cattle (unvaccinated for BHV-1) from the private farms located in Konya. The ELISA and mNT tests were used for detection of antibodies aganist the BHV-1. The Colorado reference strain of BHV-1 was used for the titration and mNT tests. The virus was propagated and titrated in the permanent Madin-Darby Bovine Kidney (MDBK) cells. Twenty-six (8.6 %) sera samples were detected positive by the ELISA applied for the detection of antibodies aganist the BHV-1. 22 (7.3 %) of totaly 301 cattle blood samples for antibodies to the BHV-1 as seropositive by the mNT. Serum Neutralisation 50 (SN) values of positive samples ranged between 1/1.7 and 1/31.6. In conclusion, the sensitivity and spesifity of tests were detected by the values obtained and their advantages were compared. For the ELISA, the sensitivity and specifity were determined as 100 % and 98 %, respectively while the corresponding values were 84 % and 100 % for the mNT.

Keywords

• BHV-1, Cattle, ELISA, Micro Neutralisation Test.

Sığırlarda Bovine Herpesvirus-1 Enfeksiyonlarının Enzyme-Linked Immunosorbent Assay ve Mikronötralizasyon Testi ile Karşılaştırmalı Tespiti

Öz

Bovine Herpesvirus-1 (BHV-1)’in neden olduğu Infectious Bovine Rhinotracheitis (IBR) tüm dünyada görülmekte ve sığırcılık endüstrisinde ciddi ekonomik kayıplara neden olmaktadır. Bu çalışma sığır kan serum örneklerinde BHV-1’e karşı gelişen antikor varlığının Enzyme-Linked Immunosorbent Assay (ELISA) ve mikro Nötralizasyon Test (mNT)’i ile tespit edilmesi ve enfeksiyonun serolojik teşhisinde kullanılan bu iki testin sensitivite ve spesifite değerlerinin belirlenmesi amaçları ile yapıldı. Çalışmada Konya’daki özel işletmelerden rastgele örnekleme ile temin edilen 301 adet sığır (BHV-1 yönünden aşılanmamış) kan örnekleri kullanıldı. Kan serum örnekleri BHV-1’e karşı oluşan antikor varlığı yönünden ELISA ve mNT ile incelendi. Titrasyon ve mNT testlerinde BHV-1’in ‘Colorado’ referens suşu kullanıldı. Virusun çoğaltılması ve titresinin belirlenmesi için Madin-Darby Bovine Kidney (MDBK) devamlı hücre kültürleri kullanıldı. BHV-1’e karşı oluşan antikor varlığı yönünden 26 adet (%8,6) kan serumu örneği ELISA ile 22 (%7,3) adedi ise mNT ile seropozitif tespit edildi. Pozitif bulunan serumların Serum Nötralizasyon 50 (SN50) değerleri 1/1,7 – 1/31,6 arasında belirlendi. Sonuç olarak, elde edilen değerlere göre testlerin sensitivite ve spesifiteleri ile avantajları karşılaştırıldı. ELISA için sensitivite %100, spesifite %98 olarak belirlenirken; mNT için sensitivite %84, spesifite ise %100 bulundu.

Anahtar Kelimeler

• BHV-1, ELISA, Mikro Nötralizasyon Test, Sığır

References

1. Ackermann M., Engels M., 2006. Pro and contra IBReradication. Vet. Microbiol., 113, 293-302.
2. Belak S., Ballagi-Pordany A., 1993. Application of the polymerase chain reaction (PCR) in veterinary diagnostic virology. Vet. Res. Commun., 17, 55
3. Bolat Y., Bulut H., Ozdarendeli A., Doymaz MZ., 19 Development of enzyme linked immunosorbent assay for detection of antibodies infectious bovine rhinotracheitis/ infectious pustular vulvovaginitis virus in cattle. F.Ü. Sağ. Bil. Derg., 10, 283-288. Bulut H., Bolat Y., Özdarendeli A., Doymaz MZ., Gürhan Sİ., 19 Infectious Bovine Rhinotracheitis (IBR) virus antijenlerinin lokalizasyonunun immunoperoksidaz boyama ile gösterimi. Tr. J. Vet. Anim. Sci., 267-271. Bulut O., Yavru S., 2004. Boğalarda Bovine Herpesvirus Tip-1 (BHV-1) enfeksiyonunun Enzyme Linked Immunosorbent Assay (ELISA), Polymerase Chain Reaction (PCR) ve Virus İzolasyonu (VI) metotları ile karşılaştırmalı teşhisi ve seroepidemiyolojisi. Eurasian J. Vet. Sci., 20, 61-70.
4. Burgu I., Akça Y., 1987. First isolation of IBR virus in Turkey. Trop. Anim. Health. Prod., 19, 56.
5. Cho HJ., Bohac JG., 1985. Sensitivity and spesificity of a enzyme-linked immunosorbent assay for detection of infectious bovine rhinotracheitis viral antibody in cattle. Can. J. Comp. Med., 49, 189-1
6. Collins JK., Butcher AC., Teramoto YA., Winston S., 19 Rapid detection of bovine herpesvirus type 1 antigens in nasal swap specimens with an antigen capture enzyme-linked immunosorbent assay. J. Clin. Microbiol., 21, 375-3 Dağalp SB., Yıldırım Y., Alkan F., 2001. Buzağılarda maternal BHV-1 antikor düzeylerinin belirlenmesi. Ankara Üniv. Vet. Fak. Derg., 48, 117-1
7. Durham PJK., Sillars HM., 1986. Evaluation of an enyzme-linked immunosorbent assay (ELISA) for serodiagnosis of infectious bovine rhinotracheitis infection with results of a preliminary survey. New Zealand Vet. J., 34, 27Edwards S., Gitao GC., 1987. Higly sensitive antigen detection procedures for the diagnosis of infectious bovine rhinotracheitis: Amplified ELISA and reverse passive haemagglutination. Vet. Microbiol., 13, 135-141.
8. Erhan M., Onar B., Csontas L., Hopkins IG., 1971. Serological survey on some virus and bedsonia diseases of cattle. Pendik Vet. Kont. Arş. Derg., 4, 55-58.

9. Fenner F., Bachman PA., Gibbs EPJ., Murphy FA., Studdert MJ., White DO., 1987. Herpesviridae. In ‘’Veterinary Virology’’, Academic Press, London.
10. Frey HR., Liess B., 1971. Vermehrungskinetik und verwendbarkeit eines stark zytopatogenen VDMD-Virusstammes für diagnostische untersuchungen mit der Mikrotiter-Methode. Zbl. Vet. Med., 18, 61-71.
11. Gupta PK., Saini M., Bandyopadhyay SK., Garg SK., 19 Bovine herpesvirus type 1 glycoprotein C expression in MDBK cells and its reactivity in enzyme-linked immunosorbent assay. Acta Virol., 42, 397-400. Ireri RG., Mirangi PK., Mbugua N., 1989. Rapid detection of bovid herpes virus 1 antigens by counter immunoelectrophoresis and agar-gel immunodiffusion. Trop. Anim. Health. Prod., 21, 50-54.
12. Kaerber G., 1964. In diagnostic procedures for virus and ricketsial disease. Public Health Ass., 3, 48
13. Meurens F., Schynts F., Keil GM., Muylkens B., Vanderplasschen A., Gallego P., Thiry E., 2004. Superinfection prevents recombination of the alphaherpesvirus bovine herpesvirus 1. J. Virol., 78, 3872-3879.
14. Murphy FA., Gibbs EPJ., Horzinek MC., Studdert MJ., 19 Herpesviridae. In ‘’Veterinary Virology’’, Eds., FA Murphy, EP Gibbs, MJ Studdert, MC Horzinek, 6 th ed., Academic Press, San Diego, California, USA, Chapter 18, 301-311. Muylkens B., Thiry J., Kirten P., Schynts F., Thiry E., 200 Bovine herpesvirus 1 infection and infectious bovine rhinotracheitis. Vet. Res., 38, 181-20 Nandi S., Kumar M., Manohar M., Chauhan RS., 200 Bovine herpes virus infections in cattle. Anim. Health Res. Rev., 10, 85-98. OIE, 2013. http:// www.oie.int/ internationalstandard-setting/ terrestrial-code/ accessonline/ [Erişim Tarihi 27.03.2013]
15. Öztürk F., Toker A., Yavru S., 1988. Konya Hayvancılık Merkez Araştırma Enstitüsü sığırlarında infectious bovine rhinotracheitis infectious pustuler vulvovaginitis (IBR/IPV) üzerine araştırma. Eurasian J. Vet. Sci., 4, 53
16. Rana SK., Kota SN., Samayam PN., Rajan S., Srinivasan VA., 2011. Use of real-time polymerase chain reaction to detect bovine herpesvirus 1 in frozen cattle and buffalo semen in India. Vet. Ital., 47, 313-322.
17. Schynts F., Baranowski E., Lemaire M., Thiry E., 19 A specific PCR to differentiate between gE negative vaccine and wildtype bovine herpesvirus type 1 strains. Vet. Microbiol., 66, 187-1 Smits CB., Van Manen C., Glas RD., De Gee AL., Dijkstrab T., Van Oirschot JT., Rijsewijk FA., 2000. Comparison of three polymerase chain reaction methods for routine detection of bovine herpesvirus 1 DNA in fresh bull sperma. J. Virol. Methods., 85, 65-73.
18. Straub OC., 1991. BHV 1 infections: Relevance and spread in Europe. Comp. Immun. Microbiol. Infect. Dis., 14, 175-186.
19. Vilcek S., Deliova I., Strojny L., Forgac O., Havran M., 19 The effect of the mode of sampling on BHV-1 detection in infected cattle by dot-blot hybridization. Vet. Microbiol., 36, 335-358. Weigler BJ., Babineau CA., Sherry B., Nasisse MP., 19 High sensitivity polymerase chain reaction assay for active and latent feline herpesvirus 1 infections in domestic cats. Vet. Rec., 29, 335-338. Wellenberg GJ., Verstraten E., Mars MH., Van Oirschot JT., 1998. Detection of bovine herpesvirus 1 glycoprotein E antibodies in individual milk samples by enzyme-linked immunosorbent assays. J. Clin. Microbiol., 36, 409-4
20. Zhou J., Lyaku J., Fredrickson RA., Kibenge FS., 1999. Improved detection of bovine herpesvirus 1 in artificially infected bovine sperma by protein amplification. J. Virol. Methods., 79, 181-189.