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感染两种不同株利什曼原虫(Leishmania)婴儿利什曼原虫的仓鼠寄生虫负担:“利什曼·多诺万单位”与实时 PCR。

Parasite burden in hamsters infected with two different strains of leishmania (Leishmania) infantum: "Leishman Donovan units" versus real-time PCR.

机构信息

Laboratório de Imunopatologia, Núcleo de Pesquisas em Ciências Biológicas/NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brasil.

出版信息

PLoS One. 2012;7(10):e47907. doi: 10.1371/journal.pone.0047907. Epub 2012 Oct 24.

Abstract

To develop and test new therapeutics and immune prophylaxis strategies for visceral leishmaniasis (VL), understanding tissue parasitism evolution after experimental infection with Leishmania infantum is important. Experimental infection in a hamster model (Mesocricetus auratus) reproduces several typical aspects of canine and human VL that are closely related to the inoculum's route. We quantified the parasitism in the liver and spleen of hamsters experimentally infected by various routes (intradermal, intraperitoneal, and intracardiac [IC]) and different strains of L. infantum (MHOM/BR/74/PP75 and Wild) and compared two different methodologies to evaluate tissue parasitism (Leishman Donovan units [LDU] and real-time qPCR). In addition, the quantification of specific total-IgG in the serum of uninfected and infected hamsters was determined by ELISA. The animals were followed for 1, 3, 6 and 9 months post-infection for survival analysis. We found that infection with the Wild strain by the IC route resulted in higher mortality. Positive antibody (IgG) responses were detected with higher peaks at 6 and 9 months in the IC group inoculated with PP75 strain. However, in animals infected with the Wild strain the IgG levels were elevated in all infected groups during all the time evaluated. We also observed by LDU analysis that the IC route lead to higher parasitism in the liver and spleen with both strains. Furthermore, qPCR showed higher sensitivity for identifying animals with low parasitic burden. In conclusion, qPCR can be useful for assessing parasitism in the spleen and liver of a hamster model infected with L. infantum independent of the route of infection, and this technique may become an essential tool for assessing parasite density in the hamster model after experimental treatment or immunization with potential vaccine candidates.

摘要

为了开发和测试内脏利什曼病(VL)的新疗法和免疫预防策略,了解利什曼原虫感染后组织寄生虫病的演变非常重要。在仓鼠模型(Mesocricetus auratus)中进行的实验感染复制了几种与犬和人类 VL 密切相关的典型方面,这些方面与接种途径有关。我们通过各种途径(皮内、腹腔内和心内[IC])和不同的利什曼原虫菌株(MHOM / BR / 74 / PP75 和野生型)对仓鼠进行了实验感染,并对两种不同的组织寄生虫病评估方法(利什曼 Donovan 单位[LDU]和实时 qPCR)进行了量化。此外,还通过 ELISA 法测定了未感染和感染仓鼠血清中的特异性总 IgG 定量。对动物进行了 1、3、6 和 9 个月的感染后生存分析。我们发现,通过 IC 途径感染野生型菌株导致更高的死亡率。在 IC 组中接种 PP75 菌株时,在 6 和 9 个月时检测到更高的 IgG 阳性抗体(IgG)反应峰值。然而,在感染野生型菌株的动物中,所有感染组在整个评估期间 IgG 水平均升高。我们还通过 LDU 分析观察到,两种菌株的 IC 途径均可导致肝和脾中的寄生虫病更高。此外,qPCR 显示出更高的敏感性,可用于识别寄生负担低的动物。总之,qPCR 可用于评估感染利什曼原虫的仓鼠模型的脾和肝中的寄生虫病,而与感染途径无关,该技术可能成为评估实验治疗或用潜在疫苗候选物免疫后仓鼠模型中寄生虫密度的重要工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd54/3480442/0ce2aae2f3f0/pone.0047907.g001.jpg

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