Chagas-Junior Adenizar D, McBride Alan J A, Athanazio Daniel A, Figueira Cláudio P, Medeiros Marco A, Reis Mitermayer G, Ko Albert I, McBride Flávia W C
Gonçalo Moniz Research Centre, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, BA, Brazil.
Department of Biointeraction, Health Sciences Institute, Federal University of Bahia, Salvador, BA, Brazil.
J Med Microbiol. 2009 Dec;58(Pt 12):1632-1637. doi: 10.1099/jmm.0.014050-0. Epub 2009 Aug 13.
In determining the efficacy of new vaccine candidates for leptospirosis, the primary end point is death and an important secondary end point is sterilizing immunity. However, evaluation of this end point is often hampered by the time-consuming demands and complexity of methods such as culture isolation (CI). In this study, we evaluated the use of an imprint (or touch preparation) method (IM) in detecting the presence of leptospires in tissues of hamsters infected with Leptospira interrogans serovar Copenhageni. In a dissemination study, compared to CI, the IM led to equal or improved detection of leptospires in kidney, liver, lung and blood samples collected post-infection and overall concordance was good (kappa=0.61). Furthermore, in an evaluation of hamsters immunized with a recombinant leptospiral protein-based vaccine candidate and subsequently challenged, the agreement between the CI and IM was very good (kappa=0.84). These findings indicate that the IM is a rapid method for the direct observation of Leptospira spp. that can be readily applied to evaluating infection in experimental animals and determining sterilizing immunity when screening potential vaccine candidates.
在确定新型钩端螺旋体疫苗候选物的疗效时，主要终点是死亡，一个重要的次要终点是杀菌免疫。然而，对这一终点的评估常常因诸如培养分离（CI）等方法耗时的要求和复杂性而受阻。在本研究中，我们评估了印记（或触片制备）法（IM）在检测感染问号钩端螺旋体哥本哈根血清型的仓鼠组织中钩端螺旋体存在情况的应用。在一项传播研究中，与CI相比，IM在感染后采集的肾脏、肝脏、肺和血液样本中对钩端螺旋体的检测效果相同或更好，总体一致性良好（kappa = 0.61）。此外，在对用基于重组钩端螺旋体蛋白的候选疫苗免疫并随后进行攻毒的仓鼠的评估中，CI和IM之间的一致性非常好（kappa = 0.84）。这些发现表明，IM是一种直接观察钩端螺旋体属的快速方法，可轻松应用于评估实验动物中的感染情况，并在筛选潜在疫苗候选物时确定杀菌免疫。
