Development of Real-Time Fluorescent PCR for Determination of Aspergillus niger

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.