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应用常规 PCR 和实时 PCR 技术检测动物及其外寄生虫中的利什曼原虫。

Detection of Leishmania infantum in animals and their ectoparasites by conventional PCR and real time PCR.

机构信息

Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation, Av. Prof. Moraes Rego s/n, Recife, PE 50670-420, Brazil.

出版信息

Exp Appl Acarol. 2013 Apr;59(4):473-81. doi: 10.1007/s10493-012-9611-4. Epub 2012 Nov 7.

Abstract

Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas.

摘要

内脏利什曼病(VL)是一种由利什曼原虫引起的寄生虫病,主要通过白蛉传播。然而,人们对其他节肢动物在 VL 传播中的作用有很多猜测。因此,本研究旨在评估巴西东北部一个 VL 流行地区猫、狗及其外寄生虫中是否存在利什曼原虫。从血液样本和外寄生虫中提取 DNA,通过针对利什曼原虫动基体 DNA 的常规 PCR(cPCR)和定量实时 PCR(qPCR)进行检测。共采集了 280 份血液样本(来自 5 只猫和 275 只狗)和 117 份来自狗的外寄生虫。动物显然健康,以前没有通过血清学或分子诊断方法进行过检测。总体而言,213 只(76.1%)动物和 51 只(43.6%)外寄生虫对利什曼原虫呈阳性,狗、猫和外寄生虫的平均寄生虫负荷分别为 795.2、31.9 和 9.1 fg。关于狗与其外寄生虫之间的阳性率,32 只(15.3%)阳性狗被阳性外寄生虫寄生。所用 PCR 方案之间的总体一致性为 59.2%,qPCR 比 cPCR 更有效;qPCR 仅对 34.1%的所有阳性样本呈阳性。大量阳性动物和外寄生虫也表明它们可以作为利什曼原虫在风险地区传播的哨兵或指标。

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