Coxiella Burnetii'nin kenelerden polimeraz zincir reaksiyonu (PCR) ve PCR-RFLP ile saptanması
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Date
2010
Journal Title
Journal ISSN
Volume Title
Publisher
Türkiye Atom Enerjisi Kurumu
Abstract
Coxiella burnetii, Q humması hastalığının etiyolojik etkeni olup, obligat intraselüler bir bateridir. Doğada yaygın olarak bulunur ve çeşitli hayvanlarda (sığır, konin, keçi) ve insanlarda Q-humması enfeksiyonunu oluşturur. C. burnetii sütlerden, kenelerden ve akut veya kronik Q humması ile enfekte insanlardan izole edilmiştir. Keneler, C. burnetii’nin rezervuarı ve birincil vektörleridir. Kırktan fazla kene türünün C. burnetii ile enfekte olduğu saptandığından keneler doğada enfeksiyonun belirleyicisi olarak görülmededirler.
Bu çalışmada, Türkiye’nin 38 ilinden toplam 2472 (1446 dişi, 1021 erkek, 5 nymph) adet kene toplandı. Bu keneler toplandığı ile, tür ve cinsiyetine 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’den toplam 56 kene toplanmış olup, bu keneler tür ve cinsiyetlerine göre 13 gruba ayrılmışlardır. Bu gruplardan 6’sı PCR sonucunda pozitif bulunmuştur. Ankara’dan toplanan toplam 160 kene ise ayni şekilde 53 gruba ayrılmış ve bu gruplardan sadece birinde PCR sonucunda pozitiflik saptanmıştır.
PCR ürünlerinin spesifisitesi restriksiyon analizi ile değerlendirildi. Pozitif PCR ürünleri Taq1 enzimi ile kesildiğinde C. burnetii Nine Mile RSA493 suşunda görüldüğü gibi yaklaşık 118, 57, 43 ve 39 bp’lik 4 bant ortaya konuldu.
Coxiella burnetii, as an obligate intracellular bacterium, is the etiologic agent of Q- fever. It is widely distrubuted in nattire and is responsible for infection in various animals (cattle, sheep, goat) and humans. C. burnetii has been isolated from milk, ticks and human patients with acute and chronic Q fever. Ticks are the principal vectors and reservoirs of C. burnetii. Since over 40 species ofticks have been found to be infected with C. burnetii, ticks can serve as indicators of infection in nature. In this study, total of 2472 ticks (1446 female, 1021 male and 5 nymphs) were collected from 38 provinces of Turkey. The ticks were gathered into groups of lto 7 ticks as to the provinces, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burnetii by using the primers CBl and CB2. The ticks collected from the province of Denizli (56 in total) were gathered into 13 groups according to the species and gender. From these groups, 6 were positive for C. burnetii. The ticks collected from Ankara province, total of 160 ticks, were Rouped into 53 as to their species and gender, and only one group was found to be positive for C. burnetii. The specificities of the PCR products were evaluated by restriction analysis. The positive PCR products were digested with the enzyme Taql 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).
Coxiella burnetii, as an obligate intracellular bacterium, is the etiologic agent of Q- fever. It is widely distrubuted in nattire and is responsible for infection in various animals (cattle, sheep, goat) and humans. C. burnetii has been isolated from milk, ticks and human patients with acute and chronic Q fever. Ticks are the principal vectors and reservoirs of C. burnetii. Since over 40 species ofticks have been found to be infected with C. burnetii, ticks can serve as indicators of infection in nature. In this study, total of 2472 ticks (1446 female, 1021 male and 5 nymphs) were collected from 38 provinces of Turkey. The ticks were gathered into groups of lto 7 ticks as to the provinces, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burnetii by using the primers CBl and CB2. The ticks collected from the province of Denizli (56 in total) were gathered into 13 groups according to the species and gender. From these groups, 6 were positive for C. burnetii. The ticks collected from Ankara province, total of 160 ticks, were Rouped into 53 as to their species and gender, and only one group was found to be positive for C. burnetii. The specificities of the PCR products were evaluated by restriction analysis. The positive PCR products were digested with the enzyme Taql 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).
Description
TAEWK D.N. 9455
Keywords
Coxiella burnetii, PCR, Tick, Kene, Q fever, Q humması
Citation
Altay, G. ...[ve arkadaşları]. (2010). Coxiella Burnetii'nin kenelerden polimeraz zincir reaksiyonu (PCR) ve PCR-RFLP ile saptanması. Ankara : Türkiye Atom Enerjisi Kurumu.