Figueiredo Mayra Araguaia Pereira, Di Santi Silvia Maria, Manrique Wilson Gómez, André Marcos Rogério, Machado Rosangela Zacarias
Laboratório de Parasitologia Animal, Curso de Medicina Veterinária, Universidade Federal de Rondônia - UNIR, Rolim de Moura, RO, Brasil.
Centro de Estudos da Malária, Superintendência de Controle de Endemias - SUCEN, São Paulo, SP, Brasil.
Rev Bras Parasitol Vet. 2018 Jul-Sep;27(3):363-376. doi: 10.1590/S1984-296120180043. Epub 2018 Jul 30.
The aim of this study was to identify Plasmodium spp. in blood samples from nonhuman primates (NHPs) in the state of Maranhão, 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.
本研究的目的是使用检测人类疟疾的经典技术和替代技术，鉴定马拉尼昂州非人类灵长类动物（NHPs）血样中的疟原虫种类。共分析了161份NHPs血样：141份来自野生动物筛查中心（CETAS）的圈养动物，20份来自私人保护区的自由生活动物。所使用的技术包括显微镜检查、快速诊断测试（RDT）、间接荧光抗体测试（IFAT）和分子技术（半巢式PCR、定量实时PCR和环介导等温扩增技术（LAMP））。还使用恶性疟原虫和伯氏疟原虫的可溶性棒状体蛋白，对两种血清学方法（斑点酶联免疫吸附测定（dot-ELISA）和间接酶联免疫吸附测定（ELISA））进行了标准化。在五只不同动物的玻片上鉴定出了疟原虫滋养体形式。通过RDT和LAMP检测，没有样本呈阳性。通过IFAT检测，有四份样本对间日疟原虫呈血清阳性。这些样本对ELISA的反应性较低。基于18S rRNA基因，使用定量PCR在34.16%（55/161）的样本中检测到疟原虫。测序后，两份样本与间日疟原虫的同一性为100%，一份样本与疟原虫属ZOOBH的同一性为97%，一份样本与恶性疟原虫的同一性为99%。PCR被证明是诊断NHPs样本中疟原虫最敏感的技术。
