黑曲霉实时荧光PCR检测方法的建立 |
| |
引用本文: | 刘金华,贺丹,史艳宇,黄宇,王丽,任常菲. 黑曲霉实时荧光PCR检测方法的建立[J]. 中国生物制品学杂志, 2009, 22(3) |
| |
作者姓名: | 刘金华 贺丹 史艳宇 黄宇 王丽 任常菲 |
| |
作者单位: | 刘金华(吉林大学基础医学院病原生物学教研室,长春130021;吉林出入境检验检疫局,长春,130062);贺丹,黄宇,王丽(吉林大学基础医学院病原生物学教研室,长春,130021);史艳宇,任常菲(吉林省产品质量监督检验院,长春,130022) |
| |
基金项目: | 吉林省科技发展计划重点项目 |
| |
摘 要: | 目的建立黑曲霉实时荧光PCR检测方法。方法对6种主要病原曲霉(黑曲霉、烟曲霉、杂色曲霉、构巢曲霉、土曲霉及黄曲霉)的GAPDH基因序列进行比对分析,选择黑曲霉特异位点设计引物和探针,对黑曲霉进行实时荧光PCR扩增,并检测该方法的灵敏度及特异性。结果该方法可检出2.78×10-10μg/ml的黑曲霉基因组DNA;对亲源关系较近的11株不同种曲霉及4株其他属临床常见的病原真菌进行实时荧光PCR检测,未发现有交叉反应。结论已建立了灵敏度高、特异性好的快速检测黑曲霉的实时荧光PCR方法。
|
关 键 词: | 黑曲霉 实时荧光PCR 特异性 灵敏度 |
Development of Real-Time Fluorescent PCR for Determination of Aspergillus niger |
| |
Abstract: | Objective To develop a real-time fluorescent PCR for determination of Aspergillus niger. Methods The GAPDH gene sequences of 6 kinds of pathogenic Aspergillus, i.e. Aspergillus niger, Aspergillus fumigatus, Aspergillus versicolor, As-pergillus nidulans, Aspergillus terreus and Aspergillus flavus, were compared, based on which primers and probes for real-time fluo-rescent PCR were designed according to the specific site of Aspergillus niger. The developed real-time fluorescent PCR was analyzed for sensitivity and specificity. Results By using the developed real-time fluorescent PCR, 2. 78 × 10-10 μg / ml of genomic DNA of Aspergillus niger was detected. Eleven Aspergillus niger isolates with close genetic relationship and 4 isolates of other common pathogenic fungi in clinic were determined by the developed method, and no cross reactions were observed. Conclusion A real-time fluorescent PCR for rapid determination of Aspergillus niger, with high specificity and sensitivity, was developed. |
| |
Keywords: | Aspergillus niger Real-time fluorescent PCR Specificity Sensitivity |
本文献已被 万方数据 等数据库收录! |
|