| 牛卵形巴贝斯虫巢式PCR诊断方法的建立及初步应用 |
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| 引用本文： | 黄国明,贾立军,薛书江,张守发.牛卵形巴贝斯虫巢式PCR诊断方法的建立及初步应用[J].兽医大学学报,2012(10):1484-1487. |
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| 作者姓名： | 黄国明 贾立军 薛书江 张守发 |
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| 作者单位： | 延边大学农学院,吉林延吉133002 |
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| 基金项目： | 吉林省自然科学基金资助项目（201115230） |
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| 摘 要： | 根据GenBank上发表的牛卵形巴贝斯虫CCTη基因序列设计合成2对巢式PCR引物,建立牛卵形巴贝斯虫巢式PCR诊断方法,对该方法的最佳反应条件进行了筛选,并进行了特异性、敏感性及临床样本检测试验。结果表明,建立的巢式PCR方法外引物扩增牛卵形巴贝斯虫基因组片段的长度为1 008bp,内引物为537bp;该方法扩增不出牛瑟氏泰勒虫、弓形虫、犬新孢子虫基因组DNA;最低检测DNA含量为16fg;通过对46份临床样本的检测,该巢式PCR较常规PCR阳性检出率高8.7%。本试验为牛卵形巴贝斯虫病的诊断提供了一种更为特异、敏感的检测技术。
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| 关 键 词： | 牛卵形巴贝斯虫 巢式PCR 诊断 |
Establishment and application of nested PCR assay for detection of Babesia ovata |
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| Authors: | HUANG Guo-ming JIA Li-jun XUE Shu-jiang ZHANG Shou-fa |
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| Institution: | (Agricultural College of Yanbian University,Yanji,Jilin 133002,China) |
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| Abstract: | In order to develop the nested-PCR diagnostic method for rapidly detection of Babesia ovata,two pairs of nested PCR primers were designed according to Babesia ovata DNA sequence.The optimum reaction conditions were screened,and sensitive,special and clinical samples test were carried out.The result showed that Babesia ovata genome DNA could be amplified by nested PCR,and Toxoplasma gondii,Theileria sergenti,Neospora canium genomic DNA could not.The minimum amount of DNA to detect is 16 fg.The detection of 46 clinical samples,indicated that the positive detection rate of nested PCR is 8.7% higher than conventional PCR.The established method of nested-PCR assay was sensitive and specific,and could be used to rapidly detect Babesia ovata. |
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| Keywords: | Babesia ovata nested PCR detection |
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