Design and development of a DNA array for rapid detection and genotyping of seven kinds of pathogenic microbes.
A DNA array for rapid detection and genotyping of the pathogenic microbes of epidemic hemorrhagic fever, tsutsugamushi disease, leptospirosis, malaria, schistosomiasis, cholera, and hemorrhagic colitis was developed. The specific and relatively conserved PCR primers and DNA probes were screened from the characteristic genes of the pathogenic microbes. The PCR or RT-PCR methods were established for amplifying and labeling the DNA fragments of the pathogenic microbes. All the probes with the same Tm value were synthesized chemically and modified with an NH2 at their 5' terminus, they were printed on glass slides for fabrication of a oligonucleotide DNA array. The developed DNA array could be used for detecting and genotyping the pathogenic microbes simultaneously, and they had a high sensitivity and specificity.
Li
Yuexi
Y
East-China Institute of Medicine and Biotechniques, Nanjing 210002, P R. China.
Tao
Kaihua
K
Jin
Huiying
H
Zhang
Jinhai
J
Cao
Min
M
Zhou
Jun
J
Tang
Yude
Y
Zhu
Jin
J
Guo
Hengbin
H
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification; D005838:Genotype; D018063:Hantaan virus / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007921:Leptospira interrogans / Q000235:genetics / Q000302:isolation & purification; D008828:Microbiological Techniques; D036103:Nanotechnology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012285:Orientia tsutsugamushi / Q000235:genetics / Q000302:isolation & purification; D010274:Parasitology / Q000379:methods*; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification; D012547:Schistosoma / Q000235:genetics / Q000302:isolation & purification; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification
D017931:DNA Primers; D015342:DNA Probes