Colorimetric detection of nucleic acid signature of shiga toxin producing Escherichia coli using gold nanoparticles.
Enterohemorrhagic E. coil (EHEC) serotype O157:H7 is one of the major pathogens, responsible for the severe disease outbreaks. EHEC causes diseases in humans through production of shiga-like toxin leading to bloody diarrhea. The toxin is encoded by stx2 gene in E. coli. The current methodology for detection of EHEC relies on fluorogenic-substrate based culture media or nucleic acid amplification based Real-Time Polymerase Chain Reaction assays that are either time consuming or need expensive instrumentation. In this study, the optical properties of gold nanoparticles (GNPs) have been exploited for detection of nucleic acid of Escherichia coli O157:H7. The stx2 gene representing EHEC signature has been targeted using the gold nanoparticle probes. Gold nanoparticles (GNPs) of 20 +/- 0.2 nm were synthesised by citrate reduction method and characterised by spectroscopy and transmission electron microscopy. The GNPs were functionalised with 19 and 22 bp of thiolated single stranded DNA complementary to target highly conserved 149 bp region of stx2 gene. Transmission Electron Microscopy revealed the hybridization, aggregation and reduction in the interparticle distances of the GNP probes in the presence of target DNA. The aggregation and the spectral shift in the plasmon band observed with 10(6) copies of target DNA indicates feasibility of a simple and quick colorimetric 'spot and read' test in contrast to amplification based detection methods.
Environmental Microbiology, Indian Institute of Toxicology Research (CSIR), Post Box 80, Mahatma Gandhi Marg, Lucknow 226001, U. P., India.
Anurag
A
Jyoti
Pratibha
P
Pandey
Surinder Pal
SP
Singh
Swatantra Kumar
SK
Jain
Rishi
R
Shanker
D001483:Base Sequence; D003124:Colorimetry; D017931:DNA Primers; D004926:Escherichia coli / Q000378:metabolism*; D006046:Gold; D053768:Metal Nanoparticles; D046529:Microscopy, Electron, Transmission; D009696:Nucleic Acids / Q000032:analysis*; D022621:Shiga Toxin / Q000096:biosynthesis*
D017931:DNA Primers; D009696:Nucleic Acids; D006046:Gold; D022621:Shiga Toxin