摘要
为建立鸡烟曲霉菌快速检测方法,根据GenBank已发表的benAfum基因序列设计1对特异性引物,建立了检测烟曲霉菌的SYBR GreenⅠ实时荧光定量PCR方法,并用该方法对烟曲霉菌分离株进行了检测。结果显示,该方法灵敏度高,对烟曲霉菌最低检出量为10 copies/L;重复性好,变异系数在0.7%~0.8%之间;特异性强,与黄曲霉、土曲霉、黑曲霉、塔宾曲霉及白色念珠菌等病原菌无交叉反应;该方法操作简单、耗时短,只需2 h即可完成整个试验过程。上述结果表明,该方法可应用于烟曲霉菌感染的快速检测。
For the purpose of setting up a convenient,discriminative and sensitive method to detect Aspegillus fumigatus in chicken,according to the ben Afum gene sequence of A.fumigatus in GenBank,a pair of special primers were designed.SYBR GreenⅠreal-time PCR for detecting A.fumigatus in chicken was established.The developed method was used to detect A.fumigatus strains.In result,the method was high sensitive with the lowest detection limit being 10 copies/L and with a good reproducibility.The variation coefficient ranged from 0.7% to 0.8%.The methed was highly discriminative without the same reactions with the other pathogenic fungi as Aspergillus flavus,Aspergillus terreus,Aspergillus niger,Aspergillus tubingensis and Candida albicans,took less time to complete the test,being 2 h.The above-mentioned results showed that the developed SYBR GreenⅠreal-time PCR could be applied to clinical detection of A.fumigatus in chicken.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第9期1135-1139,共5页
Chinese Veterinary Science
基金
河北省科技厅项目(2016D15201)
