Sığır, kanatlı ve insanlardan izole edilen mikobakterilerin moleküler tekniklerle tiplendirilmesi
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Dosyalar
Tarih
2014
Yazarlar
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Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, sığır, kanatlı ve insanlardan mikobakterilerin izolasyonu, direk klinik materyalden PZR ile tespiti, rutin teşhiste kullanılabilirliği, moleküler tiplendirilmesi ve izolatların orjinlerinin belirlenmesi amaçlandı. Toplam 81 sığır granülom ve lenf düğümü ile 9 güvercin, 2 tavuk, 1 hindi ve 1 devekuşundan doku ve organ örnekleri toplandı. Örnekler Konya'daki kesimhaneler, kanatlı yetiştiricileri ve Konya Veteriner Kontrol Enstitüsüne gönderilen tüberküloz şüpheli materyalleerden alındı. İnsan izolatları (n=50) ise, Türkiye Halk Sağlığı Kurumu Tüberküloz Laboratuvarı kültür koleksiyonundan alındı. Doku örnekleri Zielh-Neelsen ile boyandı, Lowenstein-Jensen (LJ) besiyerine ekildi ve PZR analizleri (M. tuberculosis komplex ve M. avium'un tespiti, identifikasyonu ve Restriksiyon Endonükleaz Analizleri (REA) ile tiplendirilmesi) yapıldı. Aynı zamanda, LJ'de izole edilen mikobakteri türleri de ZN ile boyandı ve PZR analizleri yapıldı. Sığır örneklerinin bakteriyoskopisinde; 25 granülom ve 43 lenf düğümü örneğinde mikobakterium şüpheli basiller gözlendi. Kültürde; 32 granülom ve 55 lenf düğümü örneğinden LJ'de mikobakterium şüpheli R tipi koloniler izole edildi. Kanatlı örneklerinden, 3 güvercin, 1 devekuşu ve 1 hindiye ait 4 dalak ve 4 karaciğer örneğinde bakteriyoskopi ile mikobakterium basilleri gözlendi. Aynı zamanda, LJ'de 4 dalak ve 5 karaciğer örneğinden mikobakteri şüpheli koloniler izole edildi. PZR yoklamalarında 28 granülom ve 47 lenf düğümü örneğinde ve LJ'de üreyen 87 şüpheli izolat M. tuberculosis complex olarak belirlendi. PZR ürünlerinin REA'sında; izolatların 70'i M. bovis ve 17'si M. bovis subsp. caprae olarak identifiye edildi. PZR ile kanatlılarda üreyen 9 izolatın tümünün M. avium, insanlara ait 50 izolatın ise M. tuberculosis olduğu gözlendi. MIRU-VNTR analizi ile MIRU-10, MIRU-16, MIRU-26, MIRU-31 ve MIRU-40 lokuslarının yüksek ayrım gücüne sahip olduğu tespit edildi. Sonuç olarak, sığırlarda M. bovis subsp. bovis ve M. bovis subsp. caprae'nin, kanatlılarda M. avium subsp. avium'un, insanlarda ise M. tuberculosis'in tüberkülozun ana etken olduğu belirlendi.
This study was aimed to isolate mycobacteria from bovine, poultry and human, to detect directly from clinical materials by PCR, usability in routin diagnosis, molecular typing and to determine origin of isolates. Samples were collected from 81 bovine granulomas and lymph nodules, tissues 9 pigeons, 2 chickens, 1 turkey and 1 ostrich. Samples from tuberculosis suspected materials were provided by slaughterhouses located in Konya Province, poultry production farmers and Veterinary Control Instıtute of Konya. In addition, human isolates (n=50) were obtained from culture collection of National Tuberculosis Reference Laboratory, Public Health Institution of Turkey. All the tissue samples were first stained by Ziehl-Neelsen, were them cultured on Lowenstein-Jensen (LJ) medium and were analysed by PCR (for detection of Mycobacterium tuberculosis complex and M. avium, identification and typing by Restriction Endonuclease Analyses). Also, mycobacteria strains isolated on LJ were stained by ZN and analysed by PCR. In microscopic examination of bovine samples; AFB results were positive in 25 granulomas and 43 lymph nodules. In culture; mycobacteria strains like-R type were isolated from 32 granulomas and 55 lymph nodules. In microscopic examination of poultry samples; AFB positivities were determined in 3 spleens and 4 livers belong to pigeon, ostrich and turkey. Also, suspected mycobacterial colonies were isolated from 4 spleens and 5 livers. M. tuberculosis complex were determined by PCR from 28 bovine granulomas and 47 lymph nodules and 87 mycobacteria isolates. According to the results of REA, a total of 70 strains were identified as M bovis and 17 were M. bovis subsp. caprae. All of the poultry isolates were detected to be M. avium, while all human isolates were M. tuberculosis. The maximal resolution of MIRU-VNTR typing was achieved using MIRU10, 16, 26, 31 and 40 loci by MIRU-VNTR analysis. In conclusion, this study shown that M. bovis subsp. bovis and M. bovis subsp. caprae in cattle, M. avium subsp. avium in poultry and M. tuberculosis in human have been found to be main agents for tuberculosis.
This study was aimed to isolate mycobacteria from bovine, poultry and human, to detect directly from clinical materials by PCR, usability in routin diagnosis, molecular typing and to determine origin of isolates. Samples were collected from 81 bovine granulomas and lymph nodules, tissues 9 pigeons, 2 chickens, 1 turkey and 1 ostrich. Samples from tuberculosis suspected materials were provided by slaughterhouses located in Konya Province, poultry production farmers and Veterinary Control Instıtute of Konya. In addition, human isolates (n=50) were obtained from culture collection of National Tuberculosis Reference Laboratory, Public Health Institution of Turkey. All the tissue samples were first stained by Ziehl-Neelsen, were them cultured on Lowenstein-Jensen (LJ) medium and were analysed by PCR (for detection of Mycobacterium tuberculosis complex and M. avium, identification and typing by Restriction Endonuclease Analyses). Also, mycobacteria strains isolated on LJ were stained by ZN and analysed by PCR. In microscopic examination of bovine samples; AFB results were positive in 25 granulomas and 43 lymph nodules. In culture; mycobacteria strains like-R type were isolated from 32 granulomas and 55 lymph nodules. In microscopic examination of poultry samples; AFB positivities were determined in 3 spleens and 4 livers belong to pigeon, ostrich and turkey. Also, suspected mycobacterial colonies were isolated from 4 spleens and 5 livers. M. tuberculosis complex were determined by PCR from 28 bovine granulomas and 47 lymph nodules and 87 mycobacteria isolates. According to the results of REA, a total of 70 strains were identified as M bovis and 17 were M. bovis subsp. caprae. All of the poultry isolates were detected to be M. avium, while all human isolates were M. tuberculosis. The maximal resolution of MIRU-VNTR typing was achieved using MIRU10, 16, 26, 31 and 40 loci by MIRU-VNTR analysis. In conclusion, this study shown that M. bovis subsp. bovis and M. bovis subsp. caprae in cattle, M. avium subsp. avium in poultry and M. tuberculosis in human have been found to be main agents for tuberculosis.
Açıklama
Anahtar Kelimeler
Kümes hayvanları, Poultry, Moleküler yapı, Molecular structure, Sığırlar, Cattles, Tüberküloz, Tuberculosis
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Sayı
Künye
Gülcü, Y. (2014). Sığır, kanatlı ve insanlardan izole edilen mikobakterilerin moleküler tekniklerle tiplendirilmesi. Selçuk Üniversitesi, Yayımlanmış doktora tezi, Konya.
