中国生物工程杂志

Objective A real-time PCR method for the detection of Aspergillus nidulans was developed. Methods Multiple nucleotide sequence alignment of 13 Aspergillus species shows high Variation in partial sequence of Glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH gene). Base on the Variation region the primers and TaqMan probes were designed for specific detecting Aspergillus nidulans specie. The specific and sensitivity of the real-time PCR was tested. Results 41 Aspergillus isolates and 12 others genus fungi were used，No cross-amplification was observed. The lowest detection limits of the real-time PCR are proved by the sensitivity testing ，4.03×10－12μg/ml of the A. nidulans DNA template can be amplified successfully. Conclusion Real-time PCR is a time-efficient，specifically method，it is a novel tool for identifying and determining the strain of A. nidulans. This method may be useful for detecting and identifying Aspergillus nidulans specie.