title: Electrochemical detection of human papillomavirus DNA type 16 using a pyrrolidinyl peptide nucleic acid probe immobilized on screen-printed carbon electrodes.
abstract: An electrochemical biosensor based on an immobilized anthraquinone-labeled pyrrolidinyl peptide nucleic acid (acpcPNA) probe was successfully developed for the selective detection of human papillomavirus (HPV) type 16 DNA. A 14-mer acpcPNA capture probe was designed to recognize a specific 14 nucleotide region of HPV type 16 L1 gene. The redox-active label anthraquinone (AQ) was covalently attached to the N-terminus of the acpcPNA probe through an amide bond. The probe was immobilized onto a chitosan-modified disposable screen-printed carbon electrode via a C-terminal lysine residue using glutaraldehyde as a cross-linking agent. Hybridization with the target DNA was studied by measuring the electrochemical signal response of the AQ label using square-wave voltammetric analysis. The calibration curve exhibited a linear range between 0.02 and 12.0 µM with a limit of detection and limit of quantitation of 4 and 14 nM, respectively. This DNA sensing platform was successfully applied to detect the HPV type 16 DNA from a PCR amplified (240 bp fragment of the L1 gene) sample derived from the HPV type 16 positive human cancer cell line (SiHa), and failed to detect the HPV-negative c33a cell line. The sensor probe exhibited very high selectivity for the complementary 14 base oligonucleotide over the non-complementary oligonucleotides with sequences derived from HPV types 18, 31 and 33. The proposed sensor provides an inexpensive tool for the early stage detection of HPV type 16, which is an important biomarker for cervical cancer.
authors.affiliation: Program in Petrochemical and Polymer Science, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
authors.forename: Sakda
authors.initials: S
authors.lastname: Jampasa
authors.affiliation: Department of Chemistry, Faculty of Science and Technology, Suan Sunandha Rajabhat University, 1 U-Thong Nok Road, Dusit, Bangkok 10300, Thailand.
authors.forename: Wanida
authors.initials: W
authors.lastname: Wonsawat
authors.affiliation: Metallurgy and Materials Science Research Institute, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
authors.forename: Nadnudda
authors.initials: N
authors.lastname: Rodthongkum
authors.affiliation: Department of Chemistry, Faculty of Science, Srinakharinwirot University, Bangkok, Thailand.
authors.forename: Weena
authors.initials: W
authors.lastname: Siangproh
authors.affiliation: Human Genetics Research Group, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Pattamawadee
authors.initials: P
authors.lastname: Yanatatsaneejit
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. Electronic address: corawon@chula.ac.th.
authors.forename: Orawon
authors.initials: O
authors.lastname: Chailapakul
mesh_terms: D000880:Anthraquinones / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation*; D002244:Carbon; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000302:isolation & purification*; D055664:Electrochemical Techniques / Q000295:instrumentation; D004566:Electrodes; D004867:Equipment Design; D052162:Human papillomavirus 16 / Q000302:isolation & purification*; D006801:Humans; D057230:Limit of Detection; D009693:Nucleic Acid Hybridization; D015341:Nucleic Acid Probes / Q000737:chemistry*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D020135:Peptide Nucleic Acids / Q000737:chemistry*
chemical_list: D000880:Anthraquinones; D004279:DNA, Viral; D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D002244:Carbon
keywords: Anthraquinone; Electrochemical detection; Human papillomavirus; Screen-printed electrode; acpcPNA