Sanderson Victoria P, Mainprize Iain L, Verzijlenberg Lisette, Khursigara Cezar M, Wills Melanie K B
G. Magnotta Lyme Disease Research Lab, Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.
Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.
Biology (Basel). 2020 Oct 29;9(11):366. doi: 10.3390/biology9110366.
Serological diagnosis of Lyme disease suffers from considerable limitations. Yet, the technique cannot currently be replaced by direct detection methods, such as bacterial culture or molecular analysis, due to their inadequate sensitivity. The low bacterial burden in vasculature and lack of consensus around blood-based isolation of the causative pathogen, , are central to this challenge. We therefore addressed methodological optimization of recovery from blood, first by analyzing existing protocols, and then by using experimentally infected human blood to identify the processing conditions and fractions that increase yield. In this proof-of-concept study, we now report two opportunities to improve recovery and detection of from clinical samples. To enhance pathogen viability and cultivability during whole blood collection, citrate anticoagulant is superior to more commonly used EDTA. Despite the widespread reliance on serum and plasma as analytes, we found that the platelet fraction of blood concentrates , providing an enriched resource for direct pathogen detection by microscopy, laboratory culture, Western blot, and PCR. The potential for platelets to serve as a reservoir for and its diagnostic targets may transform direct clinical detection of this pathogen.
莱姆病的血清学诊断存在相当大的局限性。然而，由于细菌培养或分子分析等直接检测方法灵敏度不足，目前该技术尚无法被这些方法所取代。脉管系统中细菌载量低以及在基于血液分离致病病原体方面缺乏共识，是这一挑战的核心所在。因此，我们首先通过分析现有方案，然后利用实验感染的人体血液来确定能提高回收率的处理条件和组分，从而解决了从血液中回收病原体的方法优化问题。在这项概念验证研究中，我们现在报告了两个改善从临床样本中回收和检测病原体的机会。为提高全血采集过程中病原体的活力和可培养性，柠檬酸盐抗凝剂优于更常用的乙二胺四乙酸（EDTA）。尽管血清和血浆作为分析物被广泛使用，但我们发现血液浓缩物中的血小板组分可提供丰富资源，用于通过显微镜检查、实验室培养、蛋白质印迹法和聚合酶链反应（PCR）直接检测病原体。血小板作为病原体及其诊断靶点储存库的潜力可能会改变对这种病原体的直接临床检测。
