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Development of a Real-time PCR Method to Detect Aspergillus nidulans |
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Abstract 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.
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Received: 07 July 2008
Published: 25 October 2008
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