Study On The Identification Of Candida Species In The Blood By PCR-RFLP Assay

Objective:To establish rapid methods to detect and identify Candida isolates in the blood samples by using PCR-RFLP.Methods:Biospin Fungus Genomic DNA Extraction Kit (hereinafter to be referred as Biospin) was used to isolated DNA from 50 samples which were the Candida species in the blood preparation,and compared with conventional phenol-chloroform extraction method. 20 samples which were the Candida species in the blood preparation were detected and identified by RFLP assay and compared with conventional culture method.Furthermore,detect RFLP assay's sensitivity, stability as well as specificity.Results:1 Candida albicans,Candida parapsilosis,Candida krusei can be amplified by the universal primer pair,the products were about 520bp.2 The detection limit of Candida albicans in simulated blood sample with universal primer PCR was 10CFU/ml.3 After amplification from Candida albicans,Candida parapsilosis,Candida krusei,and the products were digested with MspI,producting three unique band patterns,respectively. 4 Biospin Fungus Genomic DNA Extraction Kit,which was used to isolated DNA from Candida species is better than conventional phenol-chloroform extraction method(t=7.259,p <0.01).5 PCR-RFLP were assay detected and identified Candida species in the blood preparation is better than conventional culture method(p<0.05)and both of them have better agreement(к=0.694, p<0.01).Conclusions:PCR-RFLP assay is a rapid,sensitive and specific method to detect Candida isolates ,which is compared with conventional culture method.