Servicio de Microbiología Clínica y Enfermedades Infecciosas, Hospital General Universitario Gregorio Marañón, Madrid, Spain; Instituto de Investigación Sanitaria Gregorio Marañón, Spain; CIBER Enfermedades Respiratorias, CIBERES, Spain.
Complejo Hospitalario Torrecárdenas, Almería, Spain.
Clin Microbiol Infect. 2015 Mar;21(3):249.e1-9. doi: 10.1016/j.cmi.2014.10.003. Epub 2014 Oct 29.
Molecular epidemiology has transformed our knowledge of how tuberculosis (TB) is transmitted. Whole genome sequencing (WGS) has reached unprecedented levels of accuracy. However, it has increased technical requirements and costs, and analysis of data delays results. Our objective was to find a way to reconcile speed and ease of implementation with the high resolution of WGS. The targeted regional allele-specific oligonucleotide PCR (TRAP) assay presented here is based on allele-specific PCR targeting strain-specific single nucleotide polymorphisms, identified from WGS, and makes it possible to track actively transmitted Mycobacterium tuberculosis strains. A TRAP assay was optimized to track the most actively transmitted strains in a population in Almería, Southeast Spain, with high rates of TB. TRAP was transferred to the local laboratory where transmission was occurring. It performed well from cultured isolates and directly from sputa, enabling new secondary cases of infection from the actively transmitted strains to be detected. TRAP constitutes a fast, simple and low-cost tool that could modify surveillance of TB transmission. This pilot study could help to define a new model to survey TB transmission based on a decentralized multinodal network of local laboratories applying fast and low-cost TRAPs, which are developed by central reference centres, tailored to the specific demands of transmission at each local node.
分子流行病学改变了我们对结核病（TB）传播方式的认识。全基因组测序（WGS）已经达到了前所未有的准确性水平。然而，它增加了技术要求和成本，并且数据分析结果延迟。我们的目标是找到一种方法，将速度和易于实施与 WGS 的高分辨率相协调。这里提出的靶向区域等位基因特异性寡核苷酸 PCR（TRAP）检测法基于针对 WGS 中鉴定的菌株特异性单核苷酸多态性的等位基因特异性 PCR，可用于追踪活跃传播的结核分枝杆菌菌株。优化了 TRAP 检测法以追踪西班牙东南部阿尔梅里亚地区人群中传播率较高的最活跃传播菌株。TRAP 被转移到发生传播的当地实验室。它在培养分离物和直接从痰液中的表现良好，能够检测到活跃传播菌株的新的继发感染病例。TRAP 是一种快速、简单且低成本的工具，可以改变结核病传播的监测。这项试点研究有助于基于去中心化的多节点本地实验室网络，根据每个本地节点的传播特定需求，开发由中央参考中心定制的快速且低成本的 TRAP，来定义新的结核病传播调查模型。
