Rapid Molecular Detection of Aspergillus flavus
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摘要: 黄曲霉Aspergillus flavus是一种广泛存在的致病真菌, 早期准确检测黄曲霉并加以控制是减少黄曲霉毒素产生的有效措施。本研究利用真菌通用引物ITS1/ITS4扩增黄曲霉转录间隔区并进行克隆测序, 通过序列比对, 设计1对黄曲霉特异性引物S1/X2, 由此建立的PCR检测体系对包括黄曲霉在内的5种曲霉菌及其他17种不同的真菌、细菌基因组DNA进行扩增, 结果只有不同来源的黄曲霉菌株能扩增出334bp的特异性条带, 而其余参试菌株均无扩增产物, 其检测灵敏度在DNA水平上可达到280fg。该检测体系能从自然发病的花生或玉米组织中扩增到334bp的特异片段, 实现对黄曲霉的快速可靠检测。Abstract: Aspergillus flavus is a ubiquitous fungal pathogen in the worldwide.Early and accurate detection of A.flavus is essential to reduce the damage of Aflatoxins produced by A.flavus.Based on the difference of rDNA ITS sequences among A.flavus and other Aspergillus spp., apair of specific primers, S1/X2, was designed in this study.The primer amplified a single 334bp product from all isolates of A.flavus and that were not from other four Aspergillus species and 17 other fungi and bacteria isolates tested.The detection sensitivity was 280fg of genomic DNA.The PCR-based detection method developed here could also be used to detect A.flavus from naturally infected peanut or corn tissues.
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Key words:
- Aspergillus flavus /
- PCR technology /
- molecular detection
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