数字农科院2.0

Rapid And Sensitive Detection Of N.osema Bombycis Using Loop-Mediated I sothermal Amplification And Colorimetric Nanogold

文献类型： 外文期刊

作者： Shen, Zhongyuan;Tang, Shunming;Qi, Jingru;Shang, Ruisha;Dai, Weijiang;Chen, Hongli;Zhang, Zhilin;Tang, Shunming;Zhang, Yiling;Tang, Shunming;Shen, Zhongyuan;Zhang, Yiling;Shen, Zhongyuan;Zhang, Yiling

作者机构：

关键词： FTA card; gold nanoparticle; LAMR; microsporidia; silkworm eggs

期刊名称： SCIENCEASIA

ISSN： 1513-1874

年卷期： 2019 年 45 卷 3 期

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摘要： Salt-induced self-aggregation of gold nanoparticles (AuNP) carrying the ssDNA probes can be prevented specifically by complementary DNA. Loop-mediated isothermal amplification (LAMP) can amplify DNA rapidly. In the present study, we established a LAMP-AuNP method for detection of the microsporidium Nosema bombycis by combining the two techniques. The DNA templates of microsporidia were extracted by FTA card. The amplified product can be visually detected via hybridization at 63 degrees C for 5 min with a ssDNA-labelled nanogold probe, followed by salt-induced AuNP aggregation. The LAMP-AuNP assay needs approximately 65 min. This method can detect 10 spores/ml and is specific for N. bombycis. When it was used to detect pebrine disease of the silkworm eggs, the maximum sample size for silkworm eggs was 700, in which only one egg was infected with N. bombycis. The new LAMP-AuNP is a simple, sensitive and specific detection assay for N. bombycis.

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