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首页> 外文期刊>Berliner und Munchener Tierarztliche Wochenschrift >Comparison of different commercial DNA extraction kits to detect Toxoplasma gondii oocysts in cat faeces
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Comparison of different commercial DNA extraction kits to detect Toxoplasma gondii oocysts in cat faeces

机译：比较各种商业DNA提取试剂盒检测猫粪中弓形虫卵囊的方法

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摘要

The cat is the definitive host of Toxoplasma gondii and plays an important role in the transmission of this and other coccidian parasites, e. g. Hammondia ham-mondi, a protozoon closely related and morphologically similar to T. gondii. A number of techniques to detect T. gondii nucleic acids in feline faeces are described and several extraction kits for isolating pathogen DNA from faeces or soil are commercially available. To compare the performance of such kits with regard to isolating oocyst DNA, afeline sample that had tested negative for coccidian parasites including T. gondii and H. hammondi was spiked with 10~4,10~3, 10~2, 50 and 10 H. hammondi oocysts. Several ready-to-use stool or soil kits and an in-house method were then used to extract parasite DNA from these spiked faecal samples. Of six kits tested, two were found suitable for the detection of H. hammondi oocysts DNA by the polymerase chain reaction (PCR) in faecal samples with a detection limit of 250 oocysts per 1 g of faecal sample. These two kits revealed a similar, even slightly lower detection limit (50 oocysts per 1 g of sample) when tested with faecal samples spiked with T. gondii oocysts.
机译：猫是弓形虫的权威宿主，并且在这种和其他球虫寄生虫的传播中起着重要作用。 G。 Hammondia ham-mondi，一种与弓形虫紧密相关且在形态上相似的原生动物。描述了检测猫粪便中弓形虫核酸的许多技术，并且有几种用于从粪便或土壤中分离病原体DNA的提取试剂盒是可商购的。为了比较此类试剂盒在分离卵囊DNA方面的性能，对已检测到球虫寄生虫（包括弓形虫和哈蒙德氏菌）阴性的猫猫样品加标了10〜4、10〜3、10〜2、50和10 H hammondi卵囊。然后使用了几种现成的粪便或土壤试剂盒以及一种内部方法从这些加标的粪便样品中提取寄生虫DNA。在测试的六种试剂盒中，发现有两种适合通过粪便样品中的聚合酶链反应（PCR）检测汉莫氏酵母卵囊DNA，每1 g粪便样品的检测限为250个卵囊。当对掺有弓形虫卵囊的粪便样品进行测试时，这两种试剂盒显示出相似的甚至更低的检测限（每1 g样品50个卵囊）。

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