van Deventer A J, Goessens W H, van Belkum A, van Vliet H J, van Etten E W, Verbrugh H A
Department of Clinical Microbiology and Antimicrobial Therapy, Erasmus University School of Medicine, Rotterdam, The Netherlands.
J Clin Microbiol. 1995 Mar;33(3):625-8. doi: 10.1128/jcm.33.3.625-628.1995.
A PCR using primers aimed at the multicopy gene coding for the small subunit rRNA and resulting in the synthesis of a 180-bp fragment was evaluated for its use in diagnosing invasive candidiasis in comparison with blood culture. With the use of a C. albicans-specific probe, +/- 10 to 15 C. albicans cells are detected in 100 microliters of whole blood by Southern analysis. A DNase pretreatment was critical in the purification process of yeast DNA from whole blood. Omission of the DNase pretreatment decreased assay sensitivity 10-fold. PCR analysis of blood specimens collected from mice with invasive candidiasis is more sensitive than blood culture (100 versus 67%, respectively) at 72 h after intravenous (i.v.) inoculation with C. albicans. Furthermore, the intensity of the hybridization signals increased with the progression of infection. In contrast, multiple blood samples from gastrointestinally colonized mice were all negative by PCR, indicating that the PCR assay is also specific and may, therefore, make a positive contribution to the detection and follow-up of invasive candidiasis.
评估了一种聚合酶链反应(PCR),该反应使用针对编码小亚基核糖体RNA的多拷贝基因的引物,并合成一个180碱基对的片段,以用于与血培养相比诊断侵袭性念珠菌病。使用白色念珠菌特异性探针,通过Southern分析在100微升全血中可检测到约10至15个白色念珠菌细胞。脱氧核糖核酸酶(DNase)预处理在从全血中纯化酵母DNA的过程中至关重要。省略DNase预处理会使检测灵敏度降低10倍。在静脉内(i.v.)接种白色念珠菌72小时后,对患有侵袭性念珠菌病的小鼠采集的血液标本进行PCR分析比血培养更敏感(分别为100%对67%)。此外,杂交信号的强度随着感染的进展而增加。相比之下,胃肠道定植小鼠的多个血样经PCR检测均为阴性,表明该PCR检测具有特异性,因此可能对侵袭性念珠菌病的检测和随访有积极贡献。