title: Multiplex Paper-Based Colorimetric DNA Sensor Using Pyrrolidinyl Peptide Nucleic Acid-Induced AgNPs Aggregation for Detecting MERS-CoV, MTB, and HPV Oligonucleotides.
abstract: The development of simple fluorescent and colorimetric assays that enable point-of-care DNA and RNA detection has been a topic of significant research because of the utility of such assays in resource limited settings. The most common motifs utilize hybridization to a complementary detection strand coupled with a sensitive reporter molecule. Here, a paper-based colorimetric assay for DNA detection based on pyrrolidinyl peptide nucleic acid (acpcPNA)-induced nanoparticle aggregation is reported as an alternative to traditional colorimetric approaches. PNA probes are an attractive alternative to DNA and RNA probes because they are chemically and biologically stable, easily synthesized, and hybridize efficiently with the complementary DNA strands. The acpcPNA probe contains a single positive charge from the lysine at C-terminus and causes aggregation of citrate anion-stabilized silver nanoparticles (AgNPs) in the absence of complementary DNA. In the presence of target DNA, formation of the anionic DNA-acpcPNA duplex results in dispersion of the AgNPs as a result of electrostatic repulsion, giving rise to a detectable color change. Factors affecting the sensitivity and selectivity of this assay were investigated, including ionic strength, AgNP concentration, PNA concentration, and DNA strand mismatches. The method was used for screening of synthetic Middle East respiratory syndrome coronavirus (MERS-CoV), Mycobacterium tuberculosis (MTB), and human papillomavirus (HPV) DNA based on a colorimetric paper-based analytical device developed using the aforementioned principle. The oligonucleotide targets were detected by measuring the color change of AgNPs, giving detection limits of 1.53 (MERS-CoV), 1.27 (MTB), and 1.03 nM (HPV). The acpcPNA probe exhibited high selectivity for the complementary oligonucleotides over single-base-mismatch, two-base-mismatch, and noncomplementary DNA targets. The proposed paper-based colorimetric DNA sensor has potential to be an alternative approach for simple, rapid, sensitive, and selective DNA detection.
authors.forename: Prinjaporn
authors.initials: P
authors.lastname: Teengam
authors.affiliation: Department of Chemistry, Faculty of Science, Srinakharinwirot University , Bangkok, 10110, Thailand.
authors.forename: Weena
authors.initials: W
authors.lastname: Siangproh
authors.affiliation: Nanoelectronics and MEMS Laboratory, National Electronics and Computer Technology Center , Pathumthani 12120, Thailand.
authors.forename: Adisorn
authors.initials: A
authors.lastname: Tuantranont
authors.forename: Tirayut
authors.initials: T
authors.lastname: Vilaivan
authors.forename: Orawon
authors.initials: O
authors.lastname: Chailapakul
authors.affiliation: Departments of Chemistry and Chemical and Biological Engineering, Colorado State University , Fort Collins, Colorado 80523, United States.
authors.forename: Charles S
authors.identifier: 0000-0002-8671-7728
authors.initials: CS
authors.lastname: Henry
mesh_terms: D001483:Base Sequence; D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000032:analysis*; D004269:DNA, Bacterial / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry*; D065207:Middle East Respiratory Syndrome Coronavirus / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010209:Paper; D027383:Papillomaviridae / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D012834:Silver / Q000737:chemistry
chemical_list: D004269:DNA, Bacterial; D004279:DNA, Viral; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; D012834:Silver; D004247:DNA