期刊
CLINICAL BIOCHEMISTRY
卷 46, 期 18, 页码 1852-1856出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.clinbiochem.2013.10.013
关键词
Asymmetric capillary convective PCR; Nucleic acid on-site testing; One-step nucleic acid dipstick assay
资金
- National Science and High Technology Project [2011AA02A101]
- National High Technology Research and Development Program 863 [SQ2010AA0222525001]
- Fujian Nature Science Project [2009J06020]
- Xiamen Science and Technology Project [3502Z20111048]
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. (C) 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
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