title: Colorimetric nucleic acid testing assay for RNA virus detection based on circle-to-circle amplification of padlock probes.
abstract: We developed a molecular diagnostic method for detection of RNA virus based on padlock probes and colorimetric readout. The feasibility of our approach was demonstrated by using detection of Crimean-Congo hemorrhagic fever (CCHF) virus as a model. Compared with conventional PCR-based methods, our approach does not require advanced equipment, involves easier assay design, and has a sensitivity of 10(3) viral copies/ml. By using a cocktail of padlock probes, synthetic templates representing different viral strain variants could be detected. We analyzed 34 CCHF patient samples, and all patients were correctly diagnosed when the results were compared to those of the current real-time PCR method. This is the first time that highly specific padlock probes have been applied to detection of a highly variable target sequence typical of RNA viruses.
authors.affiliation: Department of Immunology, Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, 75185 Uppsala, Sweden.
authors.forename: Rongqin
authors.initials: R
authors.lastname: Ke
authors.forename: Anna
authors.initials: A
authors.lastname: Zorzet
authors.forename: Jenny
authors.initials: J
authors.lastname: Göransson
authors.forename: Gunnel
authors.initials: G
authors.lastname: Lindegren
authors.forename: Batool
authors.initials: B
authors.lastname: Sharifi-Mood
authors.forename: Sadegh
authors.initials: S
authors.lastname: Chinikar
authors.forename: Masoud
authors.initials: M
authors.lastname: Mardani
authors.forename: Ali
authors.initials: A
authors.lastname: Mirazimi
authors.forename: Mats
authors.initials: M
authors.lastname: Nilsson
mesh_terms: D003124:Colorimetry / Q000379:methods*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009696:Nucleic Acids / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000235:genetics*; D012327:RNA Virus Infections / Q000175:diagnosis*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D009696:Nucleic Acids; D015345:Oligonucleotide Probes