基于HSP90基因的羊吕氏泰勒虫PCR检测方法的建立与评价
文献类型: 中文期刊
作者: 翟斌涛;郑红飞;李甲;包碧波;孙万奎;王义军;王耀东;张继瑜
作者机构:
关键词: 羊吕氏泰勒虫;HSP90基因;PCR;检测方法
期刊名称:中国草食动物科学
ISSN: 2095-3887
年卷期: 2025 年
页码:
收录情况: 北大核心(2023版) ; ; 农林核心(2024版)
摘要: 吕氏泰勒虫是一种严重危害养羊业的血液原虫,建立快速、准确的检测方法对其防控至关重要。为开发一种以羊吕氏泰勒虫热休克蛋白90基因(HSP90)为新靶标的PCR检测方法,本研究根据吕氏泰勒虫HSP90基因序列设计特异性引物,通过梯度PCR优化反应条件,并对方法的敏感性、特异性和重复性进行系统评价。结果表明,本研究建立的PCR检测方法的最佳退火温度为60.5 ℃,退火时间为30秒。对该方法进行的性能评价结果表明,其敏感性高,最低可检测到32.4 pg/μL的基因组DNA(敏感性较18S rRNA靶标提高了3倍以上);特异性强,与豆状囊尾蚴、多房棘球蚴、弓形虫及球虫的DNA均无交叉反应;重复性好,在不同批次和批内试验中结果稳定。对现场采集的羊血液样品的检测结果表明,18份样品的阳性检出率为27.78%,测序结果表明扩增产物为吕氏泰勒虫。本研究首次成功建立了基于HSP90基因的羊吕氏泰勒虫PCR检测方法。该方法具有高效、灵敏、特异和可靠的优点,不仅为吕氏泰勒虫的流行病学调查和精准诊断提供了新的有效工具,而且由于HSP90在泰勒虫应激生存和致病中的关键作用,本研究也为后续探索其基因功能、开发靶向药物奠定了坚实基础。
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