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牛泰勒虫和巴贝斯虫感染的分子检测

Molecular detection of Theileria and Babesia infections in cattle.

作者信息

Altay Kursat, Aydin M Fatih, Dumanli Nazir, Aktas Munir

机构信息

Department of Parasitology, Faculty of Veterinary Medicine, University of Firat, 23119 Elazig, Turkey.

出版信息

Vet Parasitol. 2008 Dec 20;158(4):295-301. doi: 10.1016/j.vetpar.2008.09.025. Epub 2008 Oct 8.

Abstract

This study was carried out to determine the presence and distribution of tick-borne haemoprotozoan parasites (Theileria and Babesia) in apparently healthy cattle in the East Black Sea Region of Turkey. A total of 389 blood samples were collected from the animals of various ages in six provinces in the region. Prevalence of infection was determined by reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified with a set of primers for members of the genera Theileria and Babesia. Amplified PCR products were hybridized onto a membrane to which generic- and species-specific oligonucleotide probes were covalently linked. RLB hybridization identified infection in 16.19% of the samples. Blood smears were also examined microscopically for Theileria and/or Babesia spp. and 5.14% were positive. All samples shown to be positive by microscopy also tested positive with RLB assay. Two Theileria (T. annulata and T. buffeli/orientalis) and three Babesia (B. bigemina, B. major and Babesia sp.) species or genotypes were identified in the region. Babesia sp. genotype shared 99% similarity with the previously reported sequences of Babesia sp. Kashi 1, Babesia sp. Kashi 2 and Babesia sp. Kayseri 1. The most frequently found species was T. buffeli/orientalis, present in 11.56% of the samples. T. annulata was identified in five samples (1.28%). Babesia infections were less frequently detected: B. bigemina was found in three samples (0.77%), B. major in two samples (0.51%) and Babesia sp. in five samples (1.28%). A single animal infected with T. buffeli/orientalis was also infected with B. bigemina.

摘要

本研究旨在确定土耳其东黑海地区表面健康的牛体内蜱传血原虫寄生虫(泰勒虫属和巴贝斯虫属)的存在情况及分布。从该地区六个省份的不同年龄段动物身上共采集了389份血样。通过反向线印迹(RLB)分析确定感染率。使用一组针对泰勒虫属和巴贝斯虫属成员的引物扩增18S核糖体RNA(rRNA)基因的高变V4区域。将扩增的PCR产物与共价连接了通用和种特异性寡核苷酸探针的膜杂交。RLB杂交在16.19%的样本中检测到感染。还通过显微镜检查血涂片以查找泰勒虫属和/或巴贝斯虫属物种,5.14%的样本呈阳性。所有经显微镜检查显示为阳性的样本通过RLB分析也呈阳性。在该地区鉴定出两种泰勒虫(环形泰勒虫和水牛泰勒虫/东方泰勒虫)以及三种巴贝斯虫(双芽巴贝斯虫、大巴贝斯虫和巴贝斯虫属某基因型)。巴贝斯虫属某基因型与先前报道的巴贝斯虫属喀什1型、巴贝斯虫属喀什2型和巴贝斯虫属开塞利1型序列具有99%的相似性。最常见的物种是水牛泰勒虫/东方泰勒虫,存在于11.56%的样本中。在五个样本(1.28%)中鉴定出环形泰勒虫。巴贝斯虫感染的检测频率较低:在三个样本(0.77%)中发现双芽巴贝斯虫,在两个样本(0.51%)中发现大巴贝斯虫,在五个样本(1.28%)中发现巴贝斯虫属某基因型。一头感染水牛泰勒虫/东方泰勒虫的动物同时也感染了双芽巴贝斯虫。

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