Loop-mediated isothermal amplification (LAMP) assays for the species-specific detection of Eimeria that infect chickens.
Loop-mediated isothermal amplification (LAMP) assays for the species-specific detection of Eimeria that infect chickens.
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DOI：
10.1186/1746-6148-7-67
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发表时间：
2011-11-03
影响因子：
2.6
通讯作者：
Blake DP
中科院分区：
文献类型：
--
作者：
Barkway CP;Pocock RL;Vrba V;Blake DP
艾美耳球虫寄生虫可引起家禽球虫病，甚至亚临床感染也会造成经济损失。尽管存在敏感的分子检测，但感染的诊断主要依赖于传统技术，包括病变评分和粪便显微镜检查，这主要是由于成本和对专业设备的需求。尽管长期被证明有效，但这些传统技术需要时间和专业知识，可能具有高度主观性，并且可能无法诊断出亚临床疾病。认识到现代家禽生产中普遍存在的经济利润紧张以及禽球虫病对世界许多地区贫困的影响，凸显了对一组简单、敏感但具有成本效益的艾美耳球虫特异性诊断检测的需求。环介导等温扩增 (LAMP) 是一种简单、快速且相对便宜的诊断工具。在这项研究中，我们针对感染鸡的七种艾美耳球虫物种开发了一组物种特异性 LAMP 检测方法。每种检测均已被证明具有真正的物种特异性，能够检测 1 到 10 个艾美耳虫基因组，相当于少于一个成熟的裂殖体。开发一种简单的方案，用于从死后收集的组织中制备模板 DNA，无需专业实验室设备，支持这些测定在艾美耳球虫感染的常规诊断中的使用。初步现场测试支持了这一假设。一系列敏感物种特异性 LAMP 检测的开发为家禽饲养业引入了一种有价值的新的、具有成本效益的工具。
Eimeria parasites can cause the disease coccidiosis in poultry and even subclinical infection can incur economic loss. Diagnosis of infection predominantly relies on traditional techniques including lesion scoring and faecal microscopy despite the availability of sensitive molecular assays, largely due to cost and the requirement for specialist equipment. Despite longstanding proven efficacy these traditional techniques demand time and expertise, can be highly subjective and may under-diagnose subclinical disease. Recognition of the tight economic margins prevailing in modern poultry production and the impact of avian coccidiosis on poverty in many parts of the world has highlighted a requirement for a panel of straightforward and sensitive, but cost-effective, Eimeria species-specific diagnostic assays. Loop-mediated isothermal amplification (LAMP) is an uncomplicated, quick and relatively inexpensive diagnostic tool. In this study we have developed a panel of species-specific LAMP assays targeting the seven Eimeria species that infect the chicken. Each assay has been shown to be genuinely species-specific with the capacity to detect between one and ten eimerian genomes, equivalent to less than a single mature schizont. Development of a simple protocol for template DNA preparation from tissue collected post mortem with no requirement for specialist laboratory equipment supports the use of these assays in routine diagnosis of eimerian infection. Preliminary field testing supports this hypothesis. Development of a panel of sensitive species-specific LAMP assays introduces a valuable new cost-effective tool for use in poultry husbandry.
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