title: A one-step dipstick assay for the on-site detection of nucleic acid.
abstract: OBJECTIVE
  We have developed a one-step nucleic acid dipstick assay (NADA) for visually detecting polymerase chain reaction (PCR) products within 3min. "One-step" means that there were no additional procedures between amplification and detection.
  
  
  METHODS
  This method was achieved through the use of asymmetric PCR and specially designed probes with appropriate melting temperature values. We initially combined one-step NADA with asymmetric capillary convective PCR (ACCPCR), an easy and rapid nucleic acid amplification technique, to construct an on-site nucleic acid diagnostic platform.
  
  
  RESULTS
  We developed a diagnostic assay for the hepatitis B virus based on the ACCPCR-NADA platform to verify its feasibility. It exhibited an analytical sensitivity of three copies per test and a broad detection spectrum including genotype A-I. It also showed 97.9% sensitivity and 100% specificity based on the results observed using 67 serum samples with the Roche COBAS AmpliPrep/COBAS TaqMan (COBAS) system as the standard for comparison.
  
  
  CONCLUSIONS
  The results provide evidence for the feasibility of using an ACCPCR-NADA platform in practical applications, especially in on-site test.
authors.affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Disease, School of Life Science, Xiamen University, Xiamen, China.
authors.forename: Shiyin
authors.initials: S
authors.lastname: Zhang
authors.forename: Miaoge
authors.initials: M
authors.lastname: Xue
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Qingrui
authors.initials: Q
authors.lastname: Chen
authors.forename: Jieyu
authors.initials: J
authors.lastname: Chen
authors.forename: Zhantong
authors.initials: Z
authors.lastname: Wang
authors.forename: Wenbin
authors.initials: W
authors.lastname: Zhou
authors.forename: Pinghei
authors.initials: P
authors.lastname: Chen
authors.forename: Ningshao
authors.initials: N
authors.lastname: Xia
authors.forename: Shengxiang
authors.initials: S
authors.lastname: Ge
mesh_terms: D004279:DNA, Viral / Q000097:blood*; D015497:HIV-1 / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation* / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral
keywords: 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride; Asymmetric capillary convective PCR; CCPCR; COBAS; EDC; EDTA; EV71; FITC; GAM; HBV; HCV; HEV; HIV; HPV; NADA; NCBI; NIDVD; National Center for Biotechnology Information; National Institute of Diagnostics and Vaccine Development in Infectious Diseases; Nucleic acid on-site testing; One-step nucleic acid dipstick assay; PCR; PVC; Roche COBAS AmpliPrep/COBAS TaqMan; SA; TAE; Tm; capillary convective PCR; ethylenediaminetetraacetic acid; fluorescein isothiocyanate; goat anti-mouse; hepatitis B virus; hepatitis C virus; hepatitis E virus; human enterovirus 71; human immunodeficiency virus; human papillomavirus; melting temperature; nt; nucleotide; one-step nucleic acid dipstick assay; polymerase chain reaction; polyvinylchloride; streptavidin; tris-acetate-EDTA