Detection of Bacterial Pathogens by Nanogold-based Gene Chip Combining with One-time PCR with Common Primers
- VernacularTitle:纳米金基因芯片结合通用引物1次PCR法同时检测多个病原菌的研究
- Author:
Dayong GU
;
Weiping LU
;
Hua WANG
;
Yuanguo ZHOU
- Publication Type:Journal Article
- Keywords:
Nanogold;
Gene chip;
Pathogen;
16S-23S rDNA intergenic spacer region
- From:
Chinese Journal of Nosocomiology
1994;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To develop a preparation technique of sample of one-time PCR with common primers based on ribotyping which was combined with the detection system of nanogold-based gene chip to detect clinical bacterial pathogens.METHODS According to the highly conserved regions of rDNA,the common primers were designed and used to amplify each target bacterial ISRs by one-time PCR,and the specific oligonucleotide probes for each target ISRs were designed,utilized to establish the new nanogold-based gene chips.After the characteristics of the chip such as sensitivity,specificity and reliability were determined,the chip was used to detect clinical samples.RESULTS The designed common primers could amplify the 12 target bacteria successfully by one-time PCR.All selected probes were of strong specificity and great reliability.The chip had high sensitivity,specificity and reliability,reaching 50 fmol/L of detection sensibility.Clinical detection results showed the chip had a great accuracy.CONCLUSIONS Compared to multi-PCR chip detection,the detection procedure and complexity of the chip are decreased significantly,and have more practical value in clinical pathogens detection.
