基因芯片诊断常见角膜致病真菌的临床应用  

Application of gene chip technique in identification of pathogens spaces in fungal keratitis

作　　者：张英朗[1] 王丽娅[2] 李智涛 孙声桃[2] 

机构地区：[1]郑州大学第一临床学院,450052 [2]河南省眼科研究所河南省角膜病重点实验室,郑州450003 [3]河南省生物工程中心

出　　处：《眼科研究》2007年第5期379-382,共4页Chinese Ophthalmic Research

基　　金：河南省杰出人才创新基金(0321002000);河南省科技攻关计划项目(0623033000)资助

摘　　要：目的采用基因芯片技术对真菌性角膜病常见致病菌种进行临床快速检测。方法针对我国临床常见的角膜致病真菌,制备寡核苷酸探针芯片,以通用引物对170例临床采集样本进行PCR扩增,将扩增产物与寡核苷酸芯片杂交,观测相应区域的荧光显色情况,与KOH涂片法和真菌培养法鉴定结果进行统计学比较。结果170例临床角膜刮片标本,KOH涂片法检测阳性113例,阳性率为66.5%;真菌培养法阳性153例,阳性率为90.0%;芯片检测阳性147例,阳性率为86.5%,经χ2检验,芯片检测法与KOH涂片法阳性率差异有统计学意义(χ2=18.896,P=0.000),芯片检测法与真菌培养法阳性率差异无统计学意义(χ2=1.020,P=0.4);芯片法与培养法检测结果的一致性比较高度符合(Kappa值=0.80)。结论寡核苷酸探针芯片检测系统与传统的实验室检测手段比较,具有较好的特异性和灵敏度,缩短了检测时间,可对真菌性角膜病的病原菌进行快速诊断。Objective The researches on utilization of gene chip technique to identify bacteria, hepatitis virus and oncogene have been reported. However,this procedure is rarely applied in fungus identification,especially in the study of cornea fungus pathogen. The purpose of this study was to develop a technology for the rapid detection and identification of common pathogens spaces causing fungal keratitis. Methods Gene chip technique with modified oligonucleotide probes was used to detect 12 spaces of fungi in 170 clinical specimens from fungal keratitis. The chromosome DNA of these fungi was amplified and labeled using a pair of universal primers,and then the PCR products were applied to the oligonucleotide on the chip. The specific hybridization corresponded to each spaces. The outcome of gene chip test was compared with smear and cultivation. Results In 170 clinical samples from fungal keratitis,the positive numbers of gene chip test was 147 with the sensitivity 86.5% ,and those of cultivation were 153 with the sensitivity for fungi 90. 0% without significant difference between these two methods（x^2 = 1. 020, P = 0. 4）, However, significant difference was found in the sensitivity between gene chip and smear test （90.0% versus 66.5% ） （x^2= 18. 896,P =0.000）. The result of gene chip test showed a good consistency with cultivation for identification of fungi （Kappa = 0. 80）. Conclusion application of gene chip can specifically detect and identify pathogens spaces of fungal keratitis. The sensitivity of the gene chip is higher than traditional smear test .

关 键 词：真菌性角膜病 菌种 基因芯片 

分 类 号：R772[医药卫生—眼科]