title: Electrochemical paper-based peptide nucleic acid biosensor for detecting human papillomavirus.
abstract: A novel paper-based electrochemical biosensor was developed using an anthraquinone-labeled pyrrolidinyl peptide nucleic acid (acpcPNA) probe (AQ-PNA) and graphene-polyaniline (G-PANI) modified electrode to detect human papillomavirus (HPV). An inkjet printing technique was employed to prepare the paper-based G-PANI-modified working electrode. The AQ-PNA probe baring a negatively charged amino acid at the N-terminus was immobilized onto the electrode surface through electrostatic attraction. Electrochemical impedance spectroscopy (EIS) was used to verify the AQ-PNA immobilization. The paper-based electrochemical DNA biosensor was used to detect a synthetic 14-base oligonucleotide target with a sequence corresponding to human papillomavirus (HPV) type 16 DNA by measuring the electrochemical signal response of the AQ label using square-wave voltammetry before and after hybridization. It was determined that the current signal significantly decreased after the addition of target DNA. This phenomenon is explained by the rigidity of PNA-DNA duplexes, which obstructs the accessibility of electron transfer from the AQ label to the electrode surface. Under optimal conditions, the detection limit of HPV type 16 DNA was found to be 2.3 nM with a linear range of 10-200 nM. The performance of this biosensor on real DNA samples was tested with the detection of PCR-amplified DNA samples from the SiHa cell line. The new method employs an inexpensive and disposable device, which easily incinerated after use and is promising for the screening and monitoring of the amount of HPV-DNA type 16 to identify the primary stages of cervical cancer.
authors.affiliation: Program in Petrochemistry, Faculty of Science, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
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.affiliation: Department of Chemistry, Colorado State University, Fort Collins, CO, 80523, United States.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Henry
authors.affiliation: Organic Synthesis Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
authors.forename: Tirayut
authors.initials: T
authors.lastname: Vilaivan
authors.affiliation: Electrochemistry and Optical Spectroscopy Research Unit, Department of Chemistry, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand; Nanotec-CU Center of Excellence on Food and Agriculture, Bangkok, 10330, Thailand. Electronic address: corawon@chula.ac.th.
authors.forename: Orawon
authors.initials: O
authors.lastname: Chailapakul
mesh_terms: D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000302:isolation & purification*; D055664:Electrochemical Techniques; D004566:Electrodes; D052162:Human papillomavirus 16 / Q000302:isolation & purification*; D006801:Humans; D009693:Nucleic Acid Hybridization; D020135:Peptide Nucleic Acids / Q000737:chemistry*
chemical_list: D004279:DNA, Viral; D020135:Peptide Nucleic Acids
keywords: Electrochemical detection; Graphene; Human papillomavirus; Paper-based electrochemical DNA biosensor; Polyaniline; acpcPNA