DESCRIPTION OF REVERSE LINE BLOTTING (RLB) TECHNIQUE AND RLB BASED STUDIES AIMED TO DETERMINE THEILERIA AND BABESIA SPECIES IN SHEEP AND GOATS IN TURKEY

Yıl 2016, Cilt: 5 Sayı: 1, 46 - 49, 30.04.2016

Mehmet Fatih Aydın

Öz

Ticks play a role in transmission of more than 200 pathogens of medical and veterinary importance. These pathogens are present in form of single or mixed infections in the vertebrate host and the vector ticks. Microscopic and serological methods have some disadvantages such as difficulties in species discrimination and cross-reaction problems. Elimination of these disadvantages has been possible by using molecular techniques. Recently reverse line blotting (RLB) method has been used for determination of multiple infections simultaneously. RLB is also useful molecular tool to discover new species and genotypes. RLB is consist of seven steps as the stages of genus level PCR, binding species-specific probes to the Biodyne-C membrane, denaturation of PCR products, hybridization of denatured products with probes, formation of streptavidin-POD + biotin complex, realization of the light emission of this complex and evaluation the result by transferring this on a radiographic film in a dark room. This review provides detailed information about the usage of RLB in the detection of tick-borne infections.

Anahtar Kelimeler

Tick, pathogen, PCR, RLB.

REVERSE LINE BLOTTING TEKNİĞİNİN AÇIKLANMASI VE TÜRKİYE’DE KOYUN VE KEÇİLERDE THEİLERİA VE BABESİA TÜRLERİNİN BELİRLENMESİNE YÖNELİK RLB TABANLI ÇALIŞMALAR

Öz

Keneler tıbbi ve veteriner öneme sahip 200’den fazla patojenin naklinde rol oynamaktadırlar. Bu etkenler omurgalı konakta ve vektör kenelerde tek veya miks enfeksiyonlar şeklinde bulunmaktadır. Mikroskobik ve serolojik yöntemler tür ayrımında karşılaşılan zorluklar ve çapraz reaksiyon gibi dezavantajlara sahiptir. Bu dezavantajların giderilmesi moleküler tanı metotlarının kullanılmaya başlanması ile mümkün olabilmektedir. Son yıllarda, Reverse Line Blotting (RLB) metodu kullanılarak çoklu enfeksiyonların aynı zaman diliminde tespiti yapılabilmektedir. RLB metodu, yeni tür ve genotiplerin keşfedilmesini sağlayan moleküler bir tanı metodudur. RLB testi; cins düzeyinde PZR, türe özgü probların Biodyne-C membrana bağlanması, PZR ürünlerinin denatürasyonu, denatüre ürünlerin problar ile hibridizasyonu, streptavidin-POD + biotin kompleksinin oluşumu, bu kompleksin ışık emisyonunun gerçekleşmesi ve bunun karanlık odada radyografik film üzerine aktarılarak değerlendirilmesi olmak üzere yedi aşamadan oluşmaktadır. Bu derlemede kenelerle bulaşan hastalıkların teşhisinde RLB metodunun kullanımı hakkında detaylı bilgi verilmektedir.

Anahtar Kelimeler

Kene, Patojen, PZR, RLB

Kaynakça

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