摘要
为建立禽白念珠菌便捷、特异和灵敏的检测方法,根据Gen Bank已发表的白念珠菌r DNA间转录间隔区核苷酸序列设计一对目的扩增子长度为273 bp的特异引物,采用Light Cycler实时PCR(LC-PCR)检测方法,以SYBR GreenⅠ为扩增产物荧光染色剂,对禽白念珠菌疑似病例血液样本进行检测,并用临床常见的5种病原真菌对该方法的特异性进行检验。结果显示,该方法灵敏度高,白念珠菌最低检出浓度为1×101CFU/m L;特异性强,与光滑念珠菌、克柔念珠菌、热带念珠菌、近平滑念珠菌、烟曲霉等病原真菌无交叉反应;耗时短,只需2 h即可完成整个试验过程。总之,该方法可应用于禽白念珠菌早期侵染的方便、准确检测,为禽白念珠菌病的及时正确防治提供可靠的依据。
For the purpose of setting up convenient,discriminative and sensitive method to detect Candida albicans in birds,by means of Light Cycler real-time PCR technique with SYBR GreenⅠas fluorescence dye for the amplicon,we designed a pair of special primers for the 273 bp objective amplicon based on the published ITS sequences of r DNAs of Candida albicans in Gen Bank,tested the suspected case of candidiasis albicans in birds with bloodstream samples and performed specificity test of the method using 5 clinically important pathogenic fungi.The results showed that the method was high sensitive with the lowest detection as 1×10^1CFU/m L of Candida albicans,high discriminative without the same reaction in the other pathogenic fungi as Candida glabrata,Candida krusei,Candida tropicalis,Candida parapsilosis and Aspergillus fumigatus,respectively and less time consumption as long as 2 h for completing the test. In a word,this method may be used to handy and accurate detection for early infection of Candida albicans in birds and offers a reliable basis for timely and correct prevention from candidiasis albicans in birds.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第6期799-804,共6页
Chinese Veterinary Science
基金
河北省自然科学基金资助项目(C2015402157)
