Melville Lynsey, Kenyon Fiona, Javed Sajid, McElarney Iain, Demeler Janina, Skuce Philip
Vet Parasitol. 2014 Dec 15;206(3-4):308-12. doi: 10.1016/j.vetpar.2014.10.022.
A major constraint on the effective control and management of helminth parasites in livestock is the lack of rapid and reliable diagnostic tests to identify the parasite species responsible for disease and to allow informed treatment decisions to be made. In the present study, we have developed a novel DNA-based assay for the detection of Haemonchus contortus eggs in ovine faecal samples, using loop-mediated isothermal amplification (or LAMP). LAMP allows for rapid detection of H. contortus DNA under isothermal incubation conditions. The robust nature of this assay negates the need for extensive DNA extraction, allowing amplification from relatively crude samples. Preliminary results suggest that LAMP is highly specific, and does not cross-react with DNA from other common co-infecting parasites. The Haemonchus LAMP assay is also highly sensitive, exhibiting a 10 times lower detection limit than the equivalent PCR; 10(-5) ng/μl and 10(-4) ng/μl DNA, respectively, allowing detection in a faecal samples containing two Haemonchus eggs per gram. Translation of this assay onto a real-time platform provided rapid results and highlighted its potential as a quantitative assay which could inform treatment decisions in the future.
家畜体内蠕虫寄生虫有效控制和管理的一个主要限制因素是缺乏快速可靠的诊断测试，以识别导致疾病的寄生虫种类并做出明智的治疗决策。在本研究中，我们开发了一种基于DNA的新型检测方法，利用环介导等温扩增技术（LAMP）检测绵羊粪便样本中的捻转血矛线虫卵。LAMP可在等温孵育条件下快速检测捻转血矛线虫DNA。该检测方法的稳健性无需进行大量DNA提取，可从相对粗糙的样本中进行扩增。初步结果表明，LAMP具有高度特异性，不会与其他常见共感染寄生虫的DNA发生交叉反应。捻转血矛线虫LAMP检测方法也具有高度敏感性，检测限比等效PCR低10倍，分别为10(-5) ng/μl和10(-4) ng/μl DNA，能够在每克含有两个捻转血矛线虫卵的粪便样本中进行检测。将该检测方法转化为实时平台可提供快速结果，并突出了其作为定量检测方法的潜力，可为未来的治疗决策提供依据。
