Nanobiophotonics, Oral Report

EXTINCTION SPECTROSCOPY AND DYNAMIC LIGHT SCATTERING DETECTION OF DNA SEQUENCES BY USING CATIONIC GOLD NANOSPHERES

Pylaev T.E.1, Khanadeev V.A.1,Khlebtsov B.N.1, Dykman L.A.1,
Bogatyrev V.A.1,2, Khlebtsov N.G.1,2

1Institute of Biochemistry and Physiology of Plants
and Microorganisms RAS, pr. Entuziastov, 13, Saratov
2Saratov State University, ul. Astrakhanskaya 83, Saratov

ABSTRACT

We introduce a new genosensing approach employing CTAB (cetyltrimethylammonium bromide)-coated positively charged colloidal gold nanoparticles (GNPs) to detect target DNA sequences by using absorption spectroscopy and dynamic light scattering. The approach is compared with a previously reported method employing unmodified CTAB-coated gold nanorods (GNRs). Both approaches are based on the observation that whereas the addition of probe and target ssDNA to CTAB-coated particles results in particle aggregation, no aggregation is observed after addition of probe and nontarget DNA sequences. Our goal was to compare the feasibility and sensitivity of both methods. A 21-mer ssDNA from the human immunodeficiency virus type 1 HIV-1 U5 long terminal repeat (LTR) sequence and a 23-mer ssDNA from the Bacillus anthracis cryptic protein and protective antigen precursor (pagA) genes were used as ssDNA models. In the case of GNRs, unexpectedly, the colorimetric test failed with perfect cigarlike particles but could be performed with dumbbell and dog-bone rods. By contrast, our approach with cationic CTAB-coated GNPs is easy to implement and possesses excellent feasibility with retention of comparable sensitivity – a 0.1 nM concentration of target cDNA can be detected with the naked eye and 10 pM by DLS measurements. The specificity of our method is illustrated by successful DLS detection of 13 base mismatches in cDNA sequences for both DNA models. These results suggest that the cationic GNPs and DLS can be used for genosensing under optimal DNA hybridization conditions without any chemical modifications of the particle surface with ssDNA molecules and signal amplification. Finally, we discuss a more than two-three-order difference in the reported estimations of the detection sensitivity of colorimetric methods (0.1 pM to 10100 pM) to show that the existing aggregation models are inconsistent with the detection limits of about 0.1-1 pM DNA and that other explanations should be developed.

Representing author

photo

Dr. Timofey Evgen'evich Pylaev

IBPPM RAS, PhD, Researcher in Lab of Nanobiotechnology, IBPPM RAS
Saratov, Russia

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