The establishment of a gene microarray for detecting the common pathogenic fungi
10.3760/cma.j.issn.1009-9158.2011.12.016
- VernacularTitle:常见病原真菌鉴定基因芯片方法的建立
- Author:
Guanhua DENG
;
Xuan ZHENG
;
Xin ZHOU
;
Yimin HU
;
Yan XIE
;
Songmei LIU
- Publication Type:Journal Article
- Keywords:
Fungi;
Oligonucleotide array sequence analysis;
Nucleic acid hybridization
- From:
Chinese Journal of Laboratory Medicine
2011;34(12):1114-1118
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a gene microarray system for detection of clinical common pathogenic fungi.MethodsThere were 8 clinical common fungi chosen as the subjects including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilokis,Candida pseudotropicalis,Aspergillus terreus,Aspergillus flavus,Aspergillus oryzae,Aspergillus fumigatus.Universal primers,probes and specific probes for the PCR amplification and microarray preparation were designed in ITS region of the fungi genomic DNA.The PCR products amplified from those fungi's genome DNA were denatured and hybridized with the probes in gene microarray.The rapid detection of fungi was based on the investigation on the fluorescent signal intensity in the chip.The detection results of gene microarray system were verified by true positive and negative clinical samples.Results There were totally 25 positive samples identified by clinical routine microbiological methods.The 10 samples identified as bacteria positive were determined as negative without fluorescent signal by the fungi gene microarray,while the 12 samples identified as fungi positive were determined as positive with certain fungus by the fungi gene microarray.And 3 artificial Candida krusei samples were detected as fungi positive,while they were failure to be identified as certain fungus.There was no fluorescent signal in positions of the 8 fungi specific probes,but there was fluorescent signal in the position of fungi universal probe.It indicated that there were fungi in the samples but it couldn't identify the species of the fungi,because the Candida krusei wasn't included in the detection fungi list of the fungi gene microarray.ConclusionsThe fungi gene microarray established by the study could detect the common fungi in clinic rapidly and accurately.This study lays technology foundation for clinical application of gene chip.
