Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Division of Communicable Disease Control, California Department of Public Health, Richmond, California, USA.
J Clin Microbiol. 2019 Dec 23;58(1). doi: 10.1128/JCM.01106-19.
Pacific Coast tick fever is a febrile illness associated with the bite of and results from an infection due to the intracellular pathogen 364D (also known by the proposed name ""). Current molecular methods for the detection of this pathogen rely on the amplification of a conserved spotted fever group rickettsial gene (A) followed by DNA sequencing of the amplicon to identify the species. This work describes the development of a 364D-specific TaqMan assay to simplify and accelerate the detection and identification processes. The assay demonstrated a sensitivity of 1 genomic copy per 4-μl sample and is highly specific for 364D. The utility of this assay for ecological and diagnostic samples was evaluated using banked specimens collected in a single-blind manner and yielded a clinical sensitivity and specificity of 100%. In conclusion, we describe the development and evaluation of a novel TaqMan real-time PCR assay for the detection and identification of 364D suitable for ecological and diagnostic applications.
太平洋海岸蜱热是一种与蜱叮咬相关的发热疾病，由细胞内病原体 364D（也称为“”）感染引起。目前用于检测这种病原体的分子方法依赖于扩增保守的斑点热群立克次体基因（A），然后对扩增子进行 DNA 测序以鉴定物种。这项工作描述了开发一种针对 364D 的 TaqMan 检测方法，以简化和加速检测和鉴定过程。该检测方法显示出对每个 4μl 样本 1 个基因组拷贝的灵敏度，并且对 364D 具有高度特异性。使用以单盲方式收集的银行标本评估了该检测方法的实用性，其临床灵敏度和特异性均为 100%。总之，我们描述了一种新型 TaqMan 实时 PCR 检测方法的开发和评估，用于检测和鉴定适合生态和诊断应用的 364D。
