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通过聚合酶链反应检测牛的热带牛泰勒虫病

Detection of tropical bovine theileriosis by polymerase chain reaction in cattle.

作者信息

Khatoon S, Kolte S W, Kurkure N V, Chopde N A, Jahan A

机构信息

Nagpur Veterinary College, Maharashtra Animal and Fishery Sciences University (MAFSU), Nagpur, 440006 MS India.

出版信息

J Parasit Dis. 2015 Mar;39(1):53-6. doi: 10.1007/s12639-013-0270-0. Epub 2013 Mar 16.

Abstract

Tropical bovine theileriosis, a tick borne disease, caused by, Theileria annulata with marked clinical signs of pyrexia (102-105 °F), enlargement of lymphnodes etc., causes heavy economic losses in terms of high mortality and morbidity rates. Diagnosis of theileriosis is mainly based on clinical symptoms and microscopic examination of stained blood smears and lymph node biopsy smears but limitations of these methods against Theileria sp. limits the specificity. Hence, to overcome the limitations, the present study reports the detection of T. annulata in blood samples of cattle by polymerase chain reaction. The study was conducted on 155 cattle having typical clinical symptoms and blood smear after staining with Giemsa stain was examined for the presence of T. annulata in RBC. The Primer sequences were used as per d'Oliveira et al. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata and the amplification of 721 bp was done. Out of the total 155 animals, 34 were positive for T. annulata by blood smear method whereas 134 samples were positive by PCR. So diagnosis of blood samples by PCR is found to be the most sensitive and specific methodology as compared to cytological blood smear examination. The sensitivity was 23.88 % and specificity was 90.47 % of blood smear method considering PCR as gold standard and it was found that PCR is more sensitive than the conventional method of examination.

摘要

热带牛泰勒虫病是一种由环形泰勒虫引起的蜱传疾病,具有高热(102-105华氏度)、淋巴结肿大等明显临床症状,在高死亡率和发病率方面造成了巨大的经济损失。泰勒虫病的诊断主要基于临床症状以及对染色血涂片和淋巴结活检涂片的显微镜检查,但这些方法针对泰勒虫属的局限性限制了其特异性。因此,为克服这些局限性,本研究报告了通过聚合酶链反应检测牛血液样本中的环形泰勒虫。该研究对155头有典型临床症状的牛进行,用吉姆萨染色后的血涂片用于检查红细胞中环形泰勒虫的存在。引物序列按照d'Oliveira等人的方法使用。该检测采用针对环形泰勒虫编码30-kDa主要裂殖子表面抗原的基因的特异性引物,扩增出721 bp片段。在总共155只动物中,血涂片法检测到34只环形泰勒虫阳性,而PCR检测有134个样本呈阳性。因此,与细胞学血涂片检查相比,通过PCR对血液样本进行诊断是最敏感和特异的方法。以PCR为金标准,血涂片法的敏感性为23.88%,特异性为90.47%,并且发现PCR比传统检查方法更敏感。

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