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首页> 外文期刊>Revista Brasileira de Parasitologia Veterinária >Técnicas sorológicas e moleculares aplicadas na identifica??o de Plasmodium spp. em amostras de primatas n?o humanos
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Técnicas sorológicas e moleculares aplicadas na identifica??o de Plasmodium spp. em amostras de primatas n?o humanos

机译:血清学和分子技术在鉴定疟原虫中的应用。在非人类灵长类动物样本中

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The aim of this study was to identify Plasmodium spp. in blood samples from nonhuman primates (NHPs) in the state of Maranh?o, using classical and alternative techniques for examination of human malaria. A total of 161 blood samples from NHPs were analyzed: 141 from captive animals at a Wildlife Screening Center (CETAS) and 20 from free-living animals in a private reserve. The techniques used were microscopy, rapid diagnostic test (RDT), Indirect fluorescent antibody test (IFAT) and molecular techniques (semi-nested PCR, quantitative real-time PCR and LAMP). Two serological methods (dot-ELISA and indirect ELISA) were also standardized with rhoptry protein-soluble antigen of P. falciparum and P. berghei . Trophozoite forms of Plasmodium sp. were identified on slides from five different animals. No samples were positive through RDT and LAMP. Four samples were seropositive for P. malariae through IFAT. The samples showed low reactivity to ELISA. Plasmodium sp. was detected in 34.16% (55/161) of the samples using qPCR based on the 18S rRNA gene. After sequencing, two samples showed 100% identityl to P. malariae , one showed 97% identity to Plasmodium sp. ZOOBH and one showed 99% identity to P. falciparum . PCR was shown to be the most sensitive technique for diagnosing Plasmodium in NHP samples.
机译:这项研究的目的是确定疟原虫属。在Maranh?o州非人类灵长类动物(NHPs)的血液样本中使用常规和替代技术检测人类疟疾。共分析了161种NHP的血液样本:其中141种来自野生动物筛查中心(CETAS)的圈养动物,另外20种来自私人保护区的自由生活动物。使用的技术是显微镜检查,快速诊断测试(RDT),间接荧光抗体测试(IFAT)和分子技术(半嵌套式PCR,定量实时PCR和LAMP)。恶性疟原虫和伯氏疟原虫的rhoptry蛋白可溶抗原还标准化了两种血清学方法(点ELISA和间接ELISA)。滋养体形式的疟原虫在来自五种不同动物的幻灯片上被鉴定出来。通过RDT和LAMP没有样品呈阳性。四个样品通过IFAT对疟疾疟原虫呈血清阳性。样品显示出对ELISA的低反应性。疟原虫使用基于18S rRNA基因的qPCR,在34.16%(55/161)的样品中检测到了H2O。测序后,两个样品显示与疟疾假单胞菌具有100%的同一性,一个样品显示与疟原虫属种具有97%的同一性。 ZOOBH和其中一位与恶性疟原虫显示99%的同一性。 PCR被证明是诊断NHP样品中疟原虫最灵敏的技术。

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