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首页> 外文期刊>Journal of Clinical Microbiology >Comparison of Six PCR Methods Using Peripheral Blood for Detection of Canine Visceral Leishmaniasis
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Comparison of Six PCR Methods Using Peripheral Blood for Detection of Canine Visceral Leishmaniasis

机译：六种使用外周血检测犬内脏利什曼病的PCR方法的比较

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The objectives of this study were to compare the sensitivities and reliabilities of different PCR methods for the diagnosis and epidemiological study of canine visceral leishmaniasis (CVL) using dog blood. We chose to work with peripheral blood, as this type of sampling is noninvasive, straightforward, and easy to repeat. Six PCR methods were compared: three primer pairs target genomic DNA, and the other three target kinetoplast (mitochondrial) DNA. Sensitivity, specificity, reproducibility, and ease of interpretation without hybridization were evaluated for each method. The assessment was first performed using artificial samples. All methods could detect less than one parasite per reaction tube. However, the sensitivities varied among the different methods by a factor of 500 on purified cultivated parasites and by a factor of 10,000 on seeded dog blood samples (i.e., from 10 to 10?3 parasite per ml of blood for the latter). Only four methods were found sufficiently reliable for the diagnosis of CVL. They were tested on 37 dogs living in an area of endemicity and grouped according to clinical status and specific serology. Only the two methods targeting kinetoplast DNA (K13A-K13B and RV1-RV2) could detect the parasite in 100% of symptomatic infected dogs. Similarly, all seropositive dogs were found PCR positive by these methods versus 62% by the genomic-DNA-based methods. Finally, these kinetoplast-based methods proved clearly superior to the others in the detection of Leishmania in asymptomatic dogs. Our data allow the discussion of the advantages and drawbacks of highly sensitive versus moderately sensitive PCR methods in diagnosis and prevalence studies of CVL.
机译：这项研究的目的是比较不同PCR方法对犬血内脏利什曼病（CVL）的诊断和流行病学研究的敏感性和可靠性。我们选择使用外周血，因为这种类型的采样是无创，直接且易于重复的。比较了六种PCR方法：三对引物靶向基因组DNA，其他三对靶向运动质体（线粒体）DNA。每种方法均评估了灵敏度，特异性，可重复性和易于解释而无需杂交的情况。首先使用人工样品进行评估。所有方法每个反应管只能检测不到一种寄生虫。然而，在不同方法中，敏感性对纯化培养的寄生虫的影响是500倍，对种子狗血样品的敏感性是10,000（即每毫升血液中10到10 ？3 寄生虫）对于后者）。仅发现四种方法足以可靠地诊断CVL。他们对生活在流行地区的37只狗进行了测试，并根据临床状况和特定血清学进行了分组。只有两种针对运动塑料DNA的方法（K13A-K13B和RV1-RV2）才能在100％有症状感染的狗中检测到寄生虫。同样，通过这些方法发现所有血清阳性狗的PCR均为阳性，而通过基于基因组DNA的方法则为62％。最后，这些基于动质体的方法在无症状犬的利什曼原虫检测中被证明明显优于其他方法。我们的数据允许讨论在CVL的诊断和患病率研究中高敏感度与中度敏感度PCR方法的优缺点。

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