Coxiella burnetii’nin kenelerden Polimeraz Zincir Reaksiyonu ve Restriction Fragment LengthPolymorphism ile saptanması

Abstract

Coxiella burnetii’nin teşhisi amacıyla Türkiye’nin 38 ilinden toplam 2472 (1446 dişi, 1021 erkek, 5 nymph) adet kene toplandı. Keneler toplandıkları il, tür ve cinsiyetlerine göre 1-7 adedi bir araya getirilerek gruplandırıldı. Gruplar DNA ekstraksiyonu sonrasında CB1 ve CB2 primerleri kullanılarak PCR ile C. burnetii’nin varlığı yönünde incelendi. Denizli’ye ait 6 grup (Toplam 56 kene ve 13 grup) ile Ankara’ya ait bir grup (Toplam 160 kene ve 53 grup) C. burnetii yönünden pozitif bulundu. Rhipicephalus turanicus, Rhipicephalus bursa ve Hyalomma excavatum türlerinin C. burnetii taşıdıkları belirlendi. Cinsiyetin etkenin taşınmasında etkili olmadığı görüldü. PCR ürünlerinin spesifitesi restriction fragment length polimorphism (RFLP) analizi ile değerlendirildi. Pozitif PCR ürünleri TaqІ enzimi ile kesilerek, C. burnetii Nine Mile RSA493 suşunda görüldüğü gibi, 118, 57, 43 ve 39 bp’lik 4 bant ortaya konuldu

Keywords

• Coxiella burnetii
• kene
• PCR

Detection of Coxiella burnetii from ticks by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism

Abstract

For the detection of Coxiella burnetii, a total of 2472 ticks (1446 female, 1021 male and 5 nymphs) was collected from cattle and sheep of 38 provinces of Turkey. The ticks were pooled into groups of 1-7 ticks of the same province, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burnetii DNA by using the primers CB1 and CB2. Six groups from the province of Denizli (13 groups of total 56 ticks), and one group from the province of Ankara (53 groups of total 160 ticks) were found to be positive for C. burnetii. The species of Rhipicephalus turanicus, Rhipicephalus bursa and Hyalomma excavatum were found to be infected with C. burnetii. The gender was not seem to have a role in transmission of the agent. The specificities of the PCR products were evaluated by the restriction fragment length polimorphism (RFLP) analysis. The positive PCR products were digested with the enzyme TaqІ and four bands in order of 118, 57, 43 and 39 bp’s were appeared such as seen in the positive control DNA (C. burnetii Nine Mile RSA493)

Keywords

• Coxiella burnetii
• PCR
• tick

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