Solunum yolu klinik belirtileri gösteren sığırlardan izole edilen BoHV-1 izolatlarının aşı suşları ile karşılaştırılması
Yükleniyor...
Dosyalar
Tarih
2024
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bovine Herpesvirus 1 (BoHV-1), sığırlarda solunum ve genital sistemleri etkileyerek, şekillendirdiği klinik bulgulara göre, farklı isimlerle adlandırılan (IBR, IPV, IPB) enfeksiyonlara neden olur. Yetiştiricilikte, BoHV-1 ile enfekte hayvanların doğru ve hızlı bir yöntemle belirlenmesi büyük önem taşımaktadır. Bu çalışma ile solunum sistemi klinik belirtilerine sahip sığırlardan alınan örneklerde, hücre kültüründe virus izolasyonu, ELISA ile viral antijen ve Real Time PCR ile de viral genom varlığının belirlenmesi amaçlandı. Ayrıca bu projede, BoHV-1 viral genom varlığı belirlenen örneklerin moleküler düzeyde tiplendirilmesi, genomik veri tabanlarının kullanılmasıyla referans izolatlarla ve aşı suşlarıyla karşılaştırılması ve örneklenen hayvanlarda BoHV-1 seroepidemiyolojisinin belirlenmesi de hedeflenmiştir. Bu doğrultuda solunum sistemi klinik belirtili 300 sığırdan kan, nazal ve konjuktival svap örnekleri alındı. Alınan örneklerin MDBK hücre kültürlerine inokülasyonları sonucunda 6 (4'ü nazal, 2'si konjuktival svap)'sında virus izole edildi. Bunların 5'i Real Time PCR ile BoHV-1 olarak identifiye edildi. Örneklerdeki viral antijen varlığının belirlenmesi amacıyla yapılan ELISA sonucunda ise 5 (3'ü nazal, 2'si konjuktival svap) örnek pozitif olarak belirlendi. Real time PCR ile gB gen bölgesine spesifik primerler kullanılarak 19 (10 nazal, 9 konjuktival svap) örnekte BoHV-1 genom varlığı saptandı. Lökosit örnekleri (300)'nin hiçbirinde hücre kültüründe virus izolasyonu, ELISA ile viral antijen ve Real Time PCR ile de viral genom varlığı tespit edilmedi. Çalışmada elde edilen bulgulara göre, BoHV-1'in teşhisi için Real Time PCR kullanımının daha uygun olduğu belirlendi. Çalışmada, BoHV-1'e karşı antikor varlığının belirlenmesi amacıyla kan serumları ELISA ile test edildi ve %18.3 oranında seropozitiflik belirlendi. Genom varlığı belirlenen örneklerin, gC gen bölgesine göre yapılan dizi ve filogenetik analizleri sonucunda, BoHV-1.2 subtipinde olduğu belirlendi. Nükleotit sekans dizilemelerine göre, pozitif örneklerin, Genbankta yer alan referans BoHV-1 izolatları ile %98.4-99.6 oranında benzer olduğu ve aşı suşu genotiplerine %99 oranında benzerlik gösterdiği belirlendi. Sonuç olarak aşı suşlarının da içerisinde bulunduğu BoHV-1.1 ve BoHV-1.2 subtipleri arasında çok az genomik farklılık olduğu ortaya konuldu. Bu nedenle ticari olarak hazırlanmış olan ve halen kullanılan aşıların, genomik benzerlikler nedeniyle, Türkiye'de tercih edilerek kullanılabileceği belirlendi.
Bovine Herpesvirus 1 (BoHV-1) affects the respiratory and genital systems in cattle and causes infections which receive various names (IBR, IPV, IPB) according to the clinical findings. Accurate and rapid identification of BoHV-1 infected animals is a priority in animal husbandary. With this study, it was aimed to isolate the virus in cell culture and to determine the presence of viral antigen by ELISA and viral genome by Real Time PCR in samples taken from cattle with clinical symptoms of the respiratory system. Also, with this project; molecular typing of samples with the presence of BoHV-1 viral genome, comparing them with reference isolates and vaccine strains using genomic databases as well as determining the BoHV-1 seroepidemiology in sampled animals were also aimed. In this regard, blood, nasal and conjunctival swab samples were taken from 300 cattle with respiratory symptoms. As a result of inoculation of these samples into MDBK cell culture, virus was isolated in 6 (4 nasal and 2 conjunctival swabs) of them. 5 of them were identified as BoHV-1 by Real Time PCR. As a result of ELISA performed to determine the presence of viral antigen in the samples, 5 samples (3 nasal and 2 conjunctival swabs) were found to be positive. Using real-time PCR and primers specific to the gB gene region, the presence of BoHV-1 genome was detected in 19 (10 nasal, 9 conjunctival swabs) samples. There was no virus isolation in cell culture, nor was there any viral antigen detected with ELISA, and no viral genome was detected by Real Time PCR in any of the leukocyte samples (300). According to the findings of this study, it was determined that the use of Real Time PCR was more appropriate for the diagnosis of BoHV-1. In the study, blood sera were tested with ELISA to determine the presence of antibodies against BoHV-1, and a seropositivity rate of 18.3% was found. As a result of the sequence and phylogenetic analyzes performed on the gC gene region of the samples with genomic presence; they were found to be the BoHV-1.2 subtype. As a result of nucleotide sequence analysis, it was determined that the positive samples were 98.4-99.6% similar to the reference BoHV-1 isolates in Genbank and 99% similar to the vaccine strain genotypes. As a result, it was revealed that there were very few genomic differences between BoHV-1.1 and BoHV-1.2 subtypes, including the vaccine strains. For this reason, it was determined that commercially prepared and currently used vaccines could be preferable for use in Turkey due to genomic similarities.
Bovine Herpesvirus 1 (BoHV-1) affects the respiratory and genital systems in cattle and causes infections which receive various names (IBR, IPV, IPB) according to the clinical findings. Accurate and rapid identification of BoHV-1 infected animals is a priority in animal husbandary. With this study, it was aimed to isolate the virus in cell culture and to determine the presence of viral antigen by ELISA and viral genome by Real Time PCR in samples taken from cattle with clinical symptoms of the respiratory system. Also, with this project; molecular typing of samples with the presence of BoHV-1 viral genome, comparing them with reference isolates and vaccine strains using genomic databases as well as determining the BoHV-1 seroepidemiology in sampled animals were also aimed. In this regard, blood, nasal and conjunctival swab samples were taken from 300 cattle with respiratory symptoms. As a result of inoculation of these samples into MDBK cell culture, virus was isolated in 6 (4 nasal and 2 conjunctival swabs) of them. 5 of them were identified as BoHV-1 by Real Time PCR. As a result of ELISA performed to determine the presence of viral antigen in the samples, 5 samples (3 nasal and 2 conjunctival swabs) were found to be positive. Using real-time PCR and primers specific to the gB gene region, the presence of BoHV-1 genome was detected in 19 (10 nasal, 9 conjunctival swabs) samples. There was no virus isolation in cell culture, nor was there any viral antigen detected with ELISA, and no viral genome was detected by Real Time PCR in any of the leukocyte samples (300). According to the findings of this study, it was determined that the use of Real Time PCR was more appropriate for the diagnosis of BoHV-1. In the study, blood sera were tested with ELISA to determine the presence of antibodies against BoHV-1, and a seropositivity rate of 18.3% was found. As a result of the sequence and phylogenetic analyzes performed on the gC gene region of the samples with genomic presence; they were found to be the BoHV-1.2 subtype. As a result of nucleotide sequence analysis, it was determined that the positive samples were 98.4-99.6% similar to the reference BoHV-1 isolates in Genbank and 99% similar to the vaccine strain genotypes. As a result, it was revealed that there were very few genomic differences between BoHV-1.1 and BoHV-1.2 subtypes, including the vaccine strains. For this reason, it was determined that commercially prepared and currently used vaccines could be preferable for use in Turkey due to genomic similarities.
Açıklama
Anahtar Kelimeler
BoHV-1, ELISA, Real Time PCR, Sekans, Virus İzolasyonu, Sequencing, Virus Isolation
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Songül, Y. (2024). Solunum yolu klinik belirtileri gösteren sığırlardan izole edilen BoHV-1 izolatlarının aşı suşları ile karşılaştırılması. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
