APPLICATION OF PCR METHOD FOR DETECTION AND SPECIES IDENTIFICATION OF TOXOCARA SPP.

E. Kaneva, N. Tsvetkova, I. Raynova
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Abstract

The ascaridoid nematodes of dogs and cats T. canis and T. cati are with widespread distribu-tion and causative agents of a disease in human and animals named Toxocariasis. Human dis-ease has several clinical forms with different clinical manifestation such as visceral, ocular, neurotoxocariasis and covert toxocariasis. The morphological methods used to differentiate the two species, especially to identify eggs or larvae, can lead to inaccurate diagnosis. This requires the use of more reliable methods, such as PCR, for identification of Toxocara species. The aim of our research is to develop in our conditions a PCR method for species identification of Toxocara and to determine its applicability on different stages of parasites. The method used by Khademvatan et al. (2013), we performed with some modifications in different forms of Toxocara - eggs, larvae and adult parasites. We used species-specific oligonucleotide primers from the ITS2 gene sequence of the ribosomal DNA - Tcan1/NC2 for T. canis and Tcat1/NC2 for T. cati. The presence of a band with a size of 380 bp, specific for T. canis, was found for all stages of the studied parasite. The described method will allow species differentiation of Toxocariasis causative agents and improve the diagnosis of the disease, as well as determine the actual spread and reservoirs of these parasites.
PCR方法在弓形虫检测及种类鉴定中的应用。
犬和猫的蛔虫线虫是一种广泛分布的寄生虫,是人类和动物弓形虫病的病原体。人畜弓形虫病有内脏型、眼型、神经型和隐蔽性弓形虫病等多种临床表现形式。形态学方法用于区分两个物种,特别是识别卵或幼虫,可能导致不准确的诊断。这就需要使用更可靠的方法,如PCR,来鉴定弓形虫的种类。我们的研究目的是在我们的条件下建立一种弓形虫物种鉴定的PCR方法,并确定其在不同阶段寄生虫的适用性。我们采用Khademvatan等人(2013)使用的方法,对不同形式的弓形虫(卵、幼虫和成虫)进行了一些修改。我们使用了从核糖体DNA ITS2基因序列中提取的物种特异性寡核苷酸引物——犬绦虫的Tcan1/NC2和猫绦虫的Tcat1/NC2。在研究的寄生虫的所有阶段都发现了一个380 bp大小的条带,这是犬绦虫特有的。所描述的方法将允许弓形虫病病原体的物种分化,改善疾病的诊断,以及确定这些寄生虫的实际传播和宿主。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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