Sensitive and simple detection of Escherichia coli strain based on time-resolved fluorescence DNA hybridization assay.
A two-probe tandem DNA hybridization assay based on time-resolved fluorescence was employed to detect Escherichia coli strain. The amino modified capture probe was covalently immobilized on the common glass slide surface. The Eu(TTA)(3)(5-NH(2)-phen) with the characteristics of long lifetime and intense luminescence was labeled with reporter probe. The original extracted DNA samples without the purification and amplification process were directly used in the hybridization assay. The concentration of capture probe, hybridization temperature, hybridization and washing time were optimized. The detection limit is about 1.49x10(3) CFU mL(-1) E. coli cells, which is comparable to the value of most microbiology methods. The proposed method has the advantages of easy operation, satisfactory sensitivity and specificity, which can provide a promising technique for monitoring the microorganisms.
Ruan
Min
M
College of Environmental Science and Engineering, Key Laboratory of Environmental Biology and Pollution Control, Ministry of Education, Hunan University, Changsha 410082, China.
Niu
Cheng-Gang
CG
Qin
Pin-Zhu
PZ
Zeng
Guang-Ming
GM
Yang
Zhao-Hui
ZH
He
Hui
H
Huang
Jing
J
D056831:Coordination Complexes / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry*; D004269:DNA, Bacterial / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005063:Europium / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods*
D056831:Coordination Complexes; D015342:DNA Probes; D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D005063:Europium