CAJ | 학술논문

目的建立并评价一种基于AllGlo~(TM)探针的侵袭性曲霉菌病诊断技术.方法对目标基因进行克隆化并制备成标准品,用自行设计的AllGlo~(TM)探针和引物对标准品进行核酸扩增,制定定量检测标准曲线,对方法的特异性、敏感性、重复性进行评价.结果黄曲霉和炯曲霉AllGlo~(TM)荧光核酸扩增的标准曲线分别为Y=-3.003X+36.825和Y=-3.052X+38.016,其批间变异系数分别为15.60%和12.94%,表示重复性良好,敏感性达10 CFU/ml,相当于100-1000拷贝转录间隔区(ITS)2基因,与其他真菌、人类基因组及细菌无交叉阳性反应,特异性强.结论这种基于AllGlo~(TM)探针的侵袭性曲霉菌病诊断技术具有较好的重复性、特异性和较高的准确度.
목적 건립병평개일충기우AllGlo~(TM)탐침적침습성곡매균병진단기술.방법 대목표기인진행극륭화병제비성표준품,용자행설계적AllGlo~(TM)탐침화인물대표준품진행핵산확증,제정정량검측표준곡선,대방법 적특이성、민감성、중복성진행평개.결과 황곡매화형곡매AllGlo~(TM)형광핵산확증적표준곡선분별위Y=-3.003X+36.825화Y=-3.052X+38.016,기비간변이계수분별위15.60%화12.94%,표시중복성량호,민감성체10 CFU/ml,상당우100-1000고패전록간격구(ITS)2기인,여기타진균、인류기인조급세균무교차양성반응,특이성강.결론 저충기우AllGlo~(TM)탐침적침습성곡매균병진단기술구유교호적중복성、특이성화교고적준학도.
Objective To establish and evaluate a diagnostic technique based on the AllGlo~(TM) probe for the invasive aspergillosis. Methods With the self-designed AllGlo~(TM) probes and primers and the standards, two standard curves of the real-time PCR based on AllGlo~(TM) probes were established respectively for A. flaws and A. fumigatus,then its specificity, sensitivity and reproducibility were evaluated respectively. Results The findings indicated that the standard curve of A. flavus was Y = - 3. 003X + 36. 825, and A. fumigatus' was Y = - 3. 052X + 38.016, and their interassay coefficient of variation respectively were 15.60% and 12. 94% , suggesting a good reproducibility. The lowest spore concentration they could be detected was 10 CFU/ml, which equated to 100-1000 copies of internal transcribed spacer (ITS)2 genes, suggesting a good sensibility. They didn't have cross-positive reaction with other fungus, human genome and bacteria, which indicated a good specificity. Conclusion The diagnostic technique based on the AllGlo?probe for the invasive aspergillosis possessed a good sensitivity, good specificity and deadly accuracy.