Kramme Stefanie, An Le Van, Khoa Nguyen Dinh, Trin Le Van, Tannich Egbert, Rybniker Jan, Fleischer Bernhard, Drosten Christian, Panning Marcus
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
J Clin Microbiol. 2009 Mar;47(3):586-9. doi: 10.1128/JCM.00997-08. Epub 2009 Jan 14.
Scrub typhus, caused by the intracellular bacterium Orientia tsutsugamushi, is a major cause of febrile illness in the Asia/Pacific region. Here, we implemented a novel real-time PCR and determined the relation of DNA target gene concentration with serum cytokine levels. The limit of detection of the novel real-time PCR was 1,062 DNA copies per ml of EDTA whole blood. Specificity was excellent as determined on a panel of blood- and skin-borne bacteria, including Rickettsia spp. as well as healthy Vietnamese blood donors. Bacterial DNA concentrations after 9 to 12 days from symptoms onset were significantly higher than in earlier or later periods (P<0.05). Significantly higher concentrations of gamma interferon (IFN-gamma) and interleukin-10 (IL-10) occurred during the acute phase of disease (<10 days from onset) as opposed to the convalescent phase (P<0.05). No significant differences were observed between the acute and the convalescent phases for tumor necrosis factor alpha (TNF-alpha) and IL-1beta concentrations. Regression analysis of DNA concentrations and cytokine levels identified a significant positive relationship for IL-10 (P<0.0182) but not for IFN-gamma, TNF-alpha, and IL-1beta. In conclusion, proinflammatory cytokines and IL-10 were differentially related to human bacteremia. They may thus be induced by different constituents of O. tsutsugamushi. As a future prospect in a clinical diagnostic laboratory, quantitative real-time PCR may serve as a reliable tool to monitor therapy and to detect treatment failure.
恙虫病由细胞内细菌恙虫病东方体引起,是亚太地区发热性疾病的主要病因。在此,我们实施了一种新型实时聚合酶链反应(PCR),并确定了DNA靶基因浓度与血清细胞因子水平的关系。新型实时PCR的检测限为每毫升乙二胺四乙酸(EDTA)全血1062个DNA拷贝。在一组血源和皮肤源细菌(包括立克次体属以及健康的越南献血者)上进行检测,结果显示其特异性极佳。症状出现后9至12天的细菌DNA浓度显著高于早期或晚期(P<0.05)。与恢复期相比,疾病急性期(发病后<10天)γ干扰素(IFN-γ)和白细胞介素-10(IL-10)的浓度显著更高(P<0.05)。肿瘤坏死因子α(TNF-α)和IL-1β浓度在急性期和恢复期之间未观察到显著差异。DNA浓度与细胞因子水平的回归分析显示,IL-10存在显著正相关(P<0.0182),而IFN-γ、TNF-α和IL-1β则无。总之,促炎细胞因子和IL-10与人类菌血症的关系不同。因此,它们可能由恙虫病东方体的不同成分诱导产生。作为临床诊断实验室未来的展望方向,定量实时PCR可作为监测治疗和检测治疗失败的可靠工具。