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Aromatic Surfactant as Aggregating Agent for Aptamer-Gold Nanoparticle-Based Detection of Plasmodium Lactate Dehydrogenase

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Abstract

A novel ssDNA aptamer (P38), with a 40 mer random region flanked by primer-binding sites on both sides, targeting Plasmodium falciparum lactate dehydrogenase (PfLDH) has been developed through systematic evolution of ligands by exponential enrichment (SELEX), including counter SELEX against human lactate dehydrogenase A and B (hLDH A and B). The 2D structure of P38 shows the presence of three stem loops with a δG of −9.18 kcal/mol. EMSA studies on P38 complexes with the increasing concentration of PfLDH revealed a dissociation constant of 0.35 µM. P38 has been exploited for the quantitative detection of PfLDH using cationic surfactant-mediated aggregation of gold nanoparticles (16-nm diameter) as an optical probe. Among the three different cationic surfactants, characterized by different hydrocarbon tail groups, benzalkonium chloride (BCK) was found to be most efficient with a limit of detection of 281 ± 11 pM. This BCK-based approach with the novel highly selective aptamer provides simple and sensitive detection of PfLDH in the clinically relevant range.

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Abbreviations

PfLDH:

Plasmodium falciparum lactate dehydrogenase

BCK:

Benzalkonium chloride

CTAB:

Cetyltrimethyl ammonium bromide

DDAB:

Didodecyl dimethyl ammonium bromide

AuNP:

Gold nanoparticle

TEM:

Transmission electron microscopy

ELISA:

Enzyme linked immunosorbent assay

PCR:

Polymerase chain reaction

RDT:

Rapid diagnostic test

SELEX:

Systematic evolution of ligands by exponential enrichment

EMSA:

Electrophoretic mobility shift assay

hLDH A:

Human lactate dehydrogenase chain A

hLDH B:

Human lactate dehydrogenase chain B

S20,w :

Sedimentation coefficient corrected for the viscosity and density of the solvent relative to that of water at 20 °C

LOD:

Limit of detection

RT:

Room temperature

NAD:

Nicotinamide adenine dinucleotide

APAD:

3-Acetylpyridine adenine dinucleotide

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Acknowledgments

We acknowledge the financial assistance of the Department of Biotechnology, India. We thank Dr. J. Mahanta and Dr. P. K. Mohapatra from the Regional medical research centre, ICMR, Dibrugarh for providing the blood spots.

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Authors and Affiliations

  1. Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Priyamvada Jain, Babina Chakma, Naveen Kumar Singh, Sanjukta Patra & Pranab Goswami

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  1. Priyamvada Jain
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  2. Babina Chakma
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Correspondence to Pranab Goswami.

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Jain, P., Chakma, B., Singh, N.K. et al. Aromatic Surfactant as Aggregating Agent for Aptamer-Gold Nanoparticle-Based Detection of Plasmodium Lactate Dehydrogenase. Mol Biotechnol 58, 497–508 (2016). https://doi.org/10.1007/s12033-016-9946-x

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