Bergman A, Fernandez V, Holmström K O, Claesson B E B, Enroth H
Department of Clinical Microbiology, Capio Diagnostik AB, Kärnsjukhuset, Skövde, Sweden.
Eur J Clin Microbiol Infect Dis. 2007 Nov;26(11):813-8. doi: 10.1007/s10096-007-0369-2.
There is a need in the clinical microbiological laboratory for rapid and reliable methods for the universal identification of fungal pathogens. Two different regions of the rDNA gene complex, the highly polymorphic ITS1 and ITS2, were amplified using primers targeting conserved regions of the 18S, 5.8S and 28S genes. After melting-point analysis of the amplified products, the T(m) of the two PCR-products were plotted into a spot diagram where all the 14 tested, clinically relevant yeasts separated with good resolution. Real-time amplification of two separate genes, melting-point analysis and two-dimensional plotting of T(m) data can be used as a broad-range method for the identification of clinical isolates of pathogenic yeast such as Candida and Cryptococcus spp.
临床微生物实验室需要快速且可靠的方法来对真菌病原体进行通用鉴定。使用靶向18S、5.8S和28S基因保守区域的引物，扩增rDNA基因复合体的两个不同区域，即高度多态性的ITS1和ITS2。对扩增产物进行熔点分析后，将两种PCR产物的熔解温度（Tm）绘制到斑点图中，14种经测试的临床相关酵母均得到了良好分辨率的分离。对两个独立基因进行实时扩增、熔点分析以及对Tm数据进行二维绘图，可作为一种广泛应用的方法来鉴定致病性酵母（如念珠菌属和隐球菌属）的临床分离株。
