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首页> 外文期刊>Parasites Vectors >A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples
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A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples

机译：基于半自动磁捕获探针的DNA提取和实时PCR方法，适用于红狐狸（狐狸狐狸）粪便样本的echinococcus多包瑞典监测

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Background Following the first finding of Echinococcus multilocularis in Sweden in 2011, 2985 red foxes (Vulpes vulpes) were analysed by the segmental sedimentation and counting technique. This is a labour intensive method and requires handling of the whole carcass of the fox, resulting in a costly analysis. In an effort to reduce the cost of labour and sample handling, an alternative method has been developed. The method is sensitive and partially automated for detection of E. multilocularis in faecal samples. The method has been used in the Swedish E. multilocularis monitoring program for 2012–2013 on more than 2000 faecal samples. Methods We describe a new semi-automated magnetic capture probe DNA extraction method and real time hydrolysis probe polymerase chain reaction assay (MC-PCR) for the detection of E. multilocularis DNA in faecal samples from red fox. The diagnostic sensitivity was determined by validating the new method against the sedimentation and counting technique in fox samples collected in Switzerland where E. multilocularis is highly endemic. Results Of 177 foxes analysed by the sedimentation and counting technique, E. multilocularis was detected in 93 animals. Eighty-two (88%, 95% C.I 79.8-93.9) of these were positive in the MC-PCR. In foxes with more than 100 worms, the MC-PCR was positive in 44 out of 46 (95.7%) cases. The two MC-PCR negative samples originated from foxes with only immature E. multilocularis worms. In foxes with 100 worms or less, (n?=?47), 38 (80.9%) were positive in the MC-PCR. The diagnostic specificity of the MC-PCR was evaluated using fox scats collected within the Swedish screening. Of 2158 samples analysed, two were positive. This implies that the specificity is at least 99.9% (C.I. = 99.7 -100). Conclusions The MC-PCR proved to have a high sensitivity and a very high specificity. The test is partially automated but also possible to perform manually if desired. The test is well suited for nationwide E. multilocularis surveillance programs where sampling of fox scats is done to reduce the costs for sampling and where a test with a high sensitivity and a very high specificity is needed.
机译：背景技术在2011年瑞典的第一次发现echInococcus multiCularis后，通过节段性沉降和计数技术分析了2985个红狐狸（狐狸狐狸）。这是一种劳动密集型方法，需要处理狐狸的整个胴体，导致昂贵的分析。为了降低劳动力和样品处理成本，已经开发了一种替代方法。该方法对粪便样本中的核心多层检测敏感，部分自动化。该方法已在瑞典语E. 2012-2013上用于2000多个粪便样本。方法描述一种新的半自动磁捕获探针DNA提取方法和实时水解探针聚合酶链反应测定（MC-PCR），用于检测来自红狐狸的粪便样品中的E.多层样品DNA。通过验证瑞士收集的福克斯样本中的沉降和计数技术来确定诊断敏感性，其中E. MultiCularis是高度流行的。通过沉降和计数技术分析的177个狐狸的结果，E.在93只动物中检测到多层血清。八十二（88％，95％C.I 79.8-93.9）在MC-PCR中是阳性的。在具有100多个蠕虫的狐狸中，MC-PCR在46个（95.7％）的病例中有44例。两种MC-PCR阴性样品源自狐狸，其仅具有未成熟的E.多层蠕虫。在100蠕虫或更少的狐狸中（n？=Δ47），38（80.9％）在MC-PCR中阳性。使用在瑞典筛选中收集的Fox Scats评估MC-PCR的诊断特异性。分析了2158个样品，两个是阳性的。这意味着特异性至少为99.9％（C.i. = 99.7 -100）。结论MC-PCR证明具有高灵敏度和非常高的特异性。该测试是部分自动化的，但如果需要，也可以手动执行。该测试非常适合全国范围内的E.多包侦察计划，其中福克斯Scats采样以降低采样的成本，并且需要具有高灵敏度和非常高的特异性的测试。

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