Genotyping of Toxoplasma gondii isolates originated from sheep by multilocus PCR-RFLP

Abstract

The aim of the study was to determine the genotype of Toxoplasma gondii isolates obtained from sheep naturally infected. Genotyping was performed on the sheep isolates (n: 10) using multiplex multilocus nested PCR-RFLP at nine loci (SAG1, SAG2, c22-8, L358, 5’-SAG2, 3’-SAG2, SAG3, BTUB and PK1). Toxoplasma gondii isolates originated from sheep possessed multiple alleles at the same loci. The isolates (n:8) showed alleles of clonal type II and III at two loci (alt. SAG2 and SAG3), type II allele at the locus (c22-8), type I and III alleles at the locus 5'SAG2 and type I and II alleles at the locus L358, type I/III alleles at the locus 3'-SAG2. One isolate showed alleles of type I at four loci (alt. SAG2 and L358), type II allele at the locus c22-8 and type I and II alleles at the locus BTUB, type I/III alleles at the locus 3'-SAG2. One isolate showed alleles of type III at three loci (SAG1, alt. SAG2 and SAG3), type II allele at two loci (c22-8 and L358), type I/III alleles at the locus 3'-SAG2 and type I and II alleles at the locus BTUB. Genotyping may provide knowledge on infection route of T. gondii in people. To the author’s knowledge, it is the first report on genotyping of T. gondii isolates originated from sheep in Turkey. Genotyping of isolates obtained from different hosts and from various regions will contribute to achieving the important knowledge about infection routes and clonal types of T. gondii in Turkey.

Keywords

• multilocus nested PCR RFLP,Toxoplasma gondii,Genotyping,sheep,Turkey

Koyun orijinli Toxoplasma gondii izolatlarının multilokus PCR-RFLP yöntemi ile genotiplendirilmesi

Abstract

Bu çalışmada doğal enfekte koyunlardan elde edilen Toxoplasma gondii izolatlarının (n:10) genotiplendirilmesi amaçlanmıştır. Bu amaçla T. gondii izolatlarının dokuz lokusu (SAG1, SAG2, c22-8, L358, 5’-SAG2, 3’-SAG2, SAG3, BTUB and PK1) multipleks multilokus nested PCR-RFLP yöntemi ile amplifiye edilmiştir. Bunun sonucunda koyun orijinli T. gondii izolatlarının bir dizi lokusta birden çok allel barındırdığı tespit edilmiştir. Koyun izolatları (n: 8) iki lokusta (alt.SAG2 ve SAG3) tip II ve III, bir lokusta (c22-8) klonal tip II, bir lokusta (5'-SAG2) tip I ve III, bir lokusta (L358) I ve II ve bir lokusta (3'-SAG2) ise tip I/III alleli göstermiştir. Bir koyun izolatı; iki lokusta (alt.SAG2 ve L358) tip I, bir lokusta (c22-8) tip II, bir lokusta (BTUB) tip I ve II, bir lokusta (3'-SAG2) ise I/III alleli göstermiştir. Sonuncu koyun izolatı ise üç lokusta (SAG1, alt.SAG2 ve SAG3) tip III, iki lokusta (c22-8 ve L358) tip II, bir lokusta (3'-SAG2) I/III, bir lokusta (BTUB) ise tip I ve II allelleri göstermiştir. Bu çalışma Türkiye’de koyunlarda T. gondii’nin genotiplendirilmesine ilişkin ilk rapordur. Genotiplendirmeye yönelik olarak parazite arakonaklık yapan farklı hayvanlardan ve çeşitli bölgelerden elde edilen izolatlar üzerinde çalışılması Türkiye’de T. gondii’nin bulaşma yolları ve tipleri hakkında önemli bilgiler elde etmeye katkı sağlayacaktır.

Keywords

• : Genotiplendirme,koyun,multilocus nested PCR RFLP,Toxoplasma gondii,Türkiye

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