A one-step dipstick assay for the on-site detection of nucleic acid

作者：Zhang Shiyin; Xue Miaoge; Zhang Jing; Chen Qingrui; Chen Jieyu; Wang Zhantong; Zhou Wenbin; Chen Pinghei; Xia Ningshao; Ge Shengxiang^*

来源：Clinical Biochemistry, 2013, 46(18): 1852-1856.

DOI：10.1016/j.clinbiochem.2013.10.013

• Asymmetric capillary convective PCR
• Nucleic acid on-site testing
• One-step nucleic acid dipstick assay

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.

• 出版日期2013-12
• 单位厦门大学