https://scholars.lib.ntu.edu.tw/handle/123456789/448094
標題: | A one-step dipstick assay for the on-site detection of nucleic acid | 作者: | Zhang, S. Xue, M. Zhang, J. Chen, Q. Chen, J. Wang, Z. Zhou, W. Chen, P. Xia, N. Ge, S. PING-HEI CHEN |
關鍵字: | Asymmetric capillary convective PCR; Nucleic acid on-site testing; One-step nucleic acid dipstick assay | 公開日期: | 2013 | 卷: | 46 | 期: | 18 | 起(迄)頁: | 1852-1856 | 來源出版物: | Clinical Biochemistry | 摘要: | Objectives: We have developed a one-step nucleic acid dipstick assay (NADA) for visually detecting polymerase chain reaction (PCR) products within 3. min. "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. Conclusion: The results provide evidence for the feasibility of using an ACCPCR-NADA platform in practical applications, especially in on-site test. ? 2013 The Canadian Society of Clinical Chemists. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/448094 | DOI: | 10.1016/j.clinbiochem.2013.10.013 | SDG/關鍵字: | agar gel electrophoresis; article; controlled study; diagnostic test accuracy study; false negative result; genotype; Hepatitis B virus; human; limit of detection; nonhuman; nucleic acid amplification; nucleic acid analysis; one step dipstick assay; polymerase chain reaction; priority journal; sensitivity and specificity; temperature; virus load; virus strain; 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride; Asymmetric capillary convective PCR; capillary convective PCR; CCPCR; COBAS; EDC; EDTA; ethylenediaminetetraacetic acid; EV71; FITC; fluorescein isothiocyanate; GAM; goat anti-mouse; HBV; HCV; hepatitis B virus; hepatitis C virus; hepatitis E virus; HEV; HIV; HPV; human enterovirus 71; human immunodeficiency virus; human papillomavirus; melting temperature; NADA; National Center for Biotechnology Information; National Institute of Diagnostics and Vaccine Development in Infectious Diseases; NCBI; NIDVD; nt; Nucleic acid on-site testing; nucleotide; one-step nucleic acid dipstick assay; One-step nucleic acid dipstick assay; PCR; polymerase chain reaction; polyvinylchloride; PVC; Roche COBAS AmpliPrep/COBAS TaqMan; SA; streptavidin; TAE; Tm; tris-acetate-EDTA; DNA, Viral; Hepacivirus; Hepatitis B virus; HIV-1; Humans; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; Sensitivity and Specificity |
顯示於: | 機械工程學系 |
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