生物学杂志

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越冬白头鹤粪便毛细线虫卵荧光PCR分子检测方法的建立

  

  1. 安徽大学 源与环境工程学院安徽大学生物多样性与湿地生态研究所,合肥230601
  • 出版日期:2015-12-18 发布日期:2015-12-18
  • 作者简介:李博,硕士研究生,研究方向:生物多样性与保护生态学,E-mail: libo0407@126.com;
  • 基金资助:
    国家自然科学基金(31172117,3147020);安徽大学研究生科研训练强化项目目(YQH100268)

Molecular detection of Capillaria by the eggs from hooded crane faecals using real-time fluorescence PCR

  1. School of Resources and Environmental Engineering and Institute of Biodiversity and Wetland Ecology of Anhui University, Hefei 230601,China
  • Online:2015-12-18 Published:2015-12-18
  • Contact: 周立志,男,教授/博导,主要研究方向:生物多样性与保护生态学,E-mail: zhoulz@ahu.edu.cn

摘要: 寄生虫感染是濒危水鸟保护关注的热点,简便、快速、准确的寄生虫检测方法是濒危野生水鸟寄生虫学研究亟待解决的问题。为探索粪便寄生虫卵快速准确的鉴别方法,本文在越冬白头鹤粪便样品中寄生虫卵分离的基础上,通过对虫卵分别进行液氮/85℃水浴、超声破碎、裂解液消化等三种前处理,进行荧光PCR分子检测,建立粪便毛细线虫卵分子检测方法。结果显示裂解液消化法处理效果最好,此方法使虫卵经一步预处理即可满足分子检测的需要,处理方法简便而有效,检测结果稳定而敏感。此检测方法可为今后毛细线虫的分子检测提供可靠手段,也可以为濒危野生鸟类的寄生虫感染调查提供便捷的方法。

Abstract: Since the parasite infection becomes a focused issue in the research of endangered waterbirds’ conservation, it is urgent to get a simple, rapid and accurate method for parasite examin. The parasite eggs were separated from faecal samples, and treated the eggs by liquid nitrogen / 85℃ water bath, ultrasonication, and cracking fluid digestion, respectively, then detected the different effects using quantitative Realtime PCR. The method used was very well to detect Capillaria by the eggs from Hooded Crane faecals using real-time fluorescence PCR. The results showed cracking fluid digestion had the best effect, this method made the eggs to meet the need of molecular detection by one step pretreatment, providing stable and sensitive detection results with simple and efficient processes. It offered a reliable molecular detection for Capillaria, and also provided a convenient way for detection of the parasitic infection in endangered wild bird species.