A strip of lateral flow gene assay using gold nanoparticles for point-of-care diagnosis of African swine fever virus in limited environment

文献类型： 外文期刊

第一作者： Wang, Zhiying

作者： Wang, Zhiying;Yu, Wenjie;Xie, Ruibin;Yang, Shuming;Chen, Ailiang

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关键词： Recombinant polymerase amplification; Lateral flow gene assay; Tail-primer; Visual observation; In limited environment

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：3.637； 五年影响因子：3.444 ）

ISSN： 1618-2642

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收录情况： SCI

摘要： Recombinase polymerase amplification (RPA) was combined with lateral flow to develop a gold nanoparticles test strip for point-of-care diagnosis of African swine fever virus (ASFV), which is called lateral flow gene assay (LFGA). Common diagnostic techniques, including polymerase chain reaction (PCR) and immunochromatography, are time-consuming and labor-intensive, and generally require costly instruments. For improvement, this assay used tailed primers to produce DNA duplexes with a single-stranded tail at one end which can hybridize with a gold nanoparticle (AuNP)-labeled oligonucleotide detection probe. And then, biotin attached to the other end of the product bound to streptavidin, which previously fixed to the test line. Therefore, there would form a sandwich structure, and gold nanoparticles labeled on the detection probe would show a red band on the test line of strip. With the low reaction temperature (37 similar to 42 degrees C) and short reaction time (30 min), LFGA can specifically identify ASFV in blood samples infected with ASFV and classical swine fever virus (CSFV), and the LOD was 10(2) copies/mu L, which was comparable to that of agarose gel electrophoresis. In addition, blood samples infected with ASFV and CSFV were tested, and it was found that the LFGA can specifically identify ASFV DNA. In conclusion, LFGA achieves visual observation of the product after rapid RPA amplification and does not require any expensive instruments during the entire process, which is very helpful for early diagnosis of ASFV.

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