Keçeli Sema Aşkın, Dündar Devrim, Tamer Gülden Sönmez
Department of Medıcal Mıcrobıology, Kocaelı Unıversıty Hospıtal, Kocaelı, Turkey.
Mycopathologia. 2016 Feb;181(1-2):67-73. doi: 10.1007/s11046-015-9944-8. Epub 2015 Sep 23.
Candida species are generally identified by conventional methods such as germ tube or morphological appearance on corn meal agar, biochemical methods using API kits and molecular biological methods. Alternative to these methods, rapid and accurate identification methods of microorganisms called matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDİ-TOF MS) has recently been described. In this study, Candida identification results by API Candida kit, API 20C AUX kit and identifications on corn meal agar (CMA) are compared with the results obtained on Vitek-MS. All results were confirmed by sequencing internal transcribed spacer (ITS) regions of rDNA. Totally, 97 Candida strains were identified by germ tube test, CMA, API and Vitek-MS. Vitek-MS results were compatible with 74.2 % of API 20C AUX and 81.4 % of CMA results. The difference between the results of API Candida and API 20C AUX was detected. The ratio of discrepancy between Vitek-MS and API 20C AUX was 25.8 %. Candida species mostly identified as C. famata or C. tropicalis by and not compatible with API kits were identified as C. albicans by Vitek-MS. Sixteen Candida species having discrepant results with Vitek-MS, API or CMA were randomly chosen, and ITS sequence analysis was performed. The results of sequencing were compatible 56.2 % with API 20C AUX, 50 % with CMA and 93.7 % with Vitek-MS. When compared with conventional identification methods, MS results are more reliable and rapid for Candida identification. MS system may be used as routine identification method in clinical microbiology laboratories.
念珠菌属通常通过传统方法进行鉴定,如芽管试验或在玉米粉琼脂上的形态外观、使用API试剂盒的生化方法以及分子生物学方法。作为这些方法的替代方法,最近描述了一种称为基质辅助激光解吸/电离飞行时间质谱(MALDİ-TOF MS)的快速准确的微生物鉴定方法。在本研究中,将API念珠菌试剂盒、API 20C AUX试剂盒的念珠菌鉴定结果以及在玉米粉琼脂(CMA)上的鉴定结果与Vitek-MS获得的结果进行了比较。所有结果均通过对rDNA的内部转录间隔区(ITS)进行测序来确认。总共通过芽管试验、CMA、API和Vitek-MS鉴定了97株念珠菌菌株。Vitek-MS的结果与API 20C AUX的74.2%以及CMA的81.4%的结果相符。检测到API念珠菌和API 20C AUX结果之间的差异。Vitek-MS与API 20C AUX之间的差异率为25.8%。通过API试剂盒鉴定为近平滑念珠菌或热带念珠菌且与API试剂盒不相符的念珠菌属,通过Vitek-MS鉴定为白色念珠菌。随机选择了16株与Vitek-MS、API或CMA结果存在差异的念珠菌属进行ITS序列分析。测序结果与API 20C AUX的相符率为56.2%,与CMA的相符率为50%,与Vitek-MS的相符率为93.7%。与传统鉴定方法相比,质谱结果在念珠菌鉴定方面更可靠、更快速。质谱系统可作为临床微生物实验室的常规鉴定方法。