He Zheng-Xin, Zhao Hui-Hai, Wang Fu-Kun
Department of Clinical Laboratory, The 980th Hospital of PLA Joint Logistical Support Force (Bethune International Peace Hospital), 398 Zhongshan Road, Shijiazhuang, Hebei, 050082, People's Republic of China.
Open Life Sci. 2020 Sep 6;15(1):677-682. doi: 10.1515/biol-2020-0075. eCollection 2020.
Invasive candidiasis is a major challenge to clinical medicine today. However, traditional fungal diagnostic techniques and empirical treatments have shown great limitations. Although efforts are necessarily needed in methodology standardization and multicenter validation, polymerase chain reaction (PCR) is a very promising assay in detecting fungal pathogens. Using a "heat-shock" DNA preparation method, a rapid and simple PCR protocol for quantification of the () ribosomal DNA was established. The PCR assay could detect DNA as low as 10 CFU/mL in samples prepared by the heat-shock protocol, without any cross-reaction with DNA prepared from other spp. and bacterial pathogens. For simulated blood samples, the PCR test sensitivity of whole blood samples was better than that of plasma and blood cells. In the systemic candidiasis murine model, detectable DNA was only observed within 24 h after SC5314 injection, which is much shorter than that observed in the kidney.
侵袭性念珠菌病是当今临床医学面临的一项重大挑战。然而，传统的真菌诊断技术和经验性治疗已显示出极大的局限性。尽管在方法标准化和多中心验证方面必然需要付出努力，但聚合酶链反应（PCR）在检测真菌病原体方面是一种非常有前景的检测方法。采用“热休克”DNA制备方法，建立了一种快速简便的用于定量（）核糖体DNA的PCR方案。该PCR检测方法能够在通过热休克方案制备的样品中检测到低至10 CFU/mL的DNA，且与从其他（）菌种和细菌病原体制备的DNA无任何交叉反应。对于模拟血样，全血样本的PCR检测灵敏度优于血浆和血细胞。在系统性念珠菌病小鼠模型中，仅在注射SC5314后24小时内观察到可检测的DNA，这比在肾脏中观察到的时间要短得多。
