Betaine-assisted recombinase polymerase assay for rapid hepatitis B virus detection,Biotechnology and Applied Biochemistry

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Betaine-assisted recombinase polymerase assay for rapid hepatitis B virus detection
Biotechnology and Applied Biochemistry ( IF 3.2 ) Pub Date : 2020-05-10 , DOI: 10.1002/bab.1940
Ting-Ting Yi _{1,

2} , Han-Yun Zhang ₃ , Hua Liang ₄ , Guo-Zhong Gong ₅ , Yan Cai _{2,

6}

Affiliation

Hepatitis B virus (HBV) is a worldwide epidemic pathogen that causes hepatitis B. On-site screening the HBV infection is of critical importance for preventing and diagnosing HBV infection. In this paper, a simple, visual, and rapid method for on-site detection of HBV-DNA has been developed. This method is based on betaine-assisted recombinase polymerase assay and followed with naked-eye detection via lateral flow assay (BRPA-LF). Result show that nonspecific amplification is prone to occur in recombinase polymerase amplification (RPA) if the assay was performed with serum sample without purification. This problem has been addressed by adding 0.8 M of betaine to the RPA reactions. It was demonstrated that BRPA-LF can detect 1,000 copies of HBV-DNA in 50 μL mixture, and achieved 90% sensitivity and 100% specificity for serum sample detection. These results demonstrated that BRPA-LF can resist serum interference and has great potential for on-site screening of HBV infection.

中文翻译：

用于快速乙型肝炎病毒检测的甜菜碱辅助重组酶聚合酶测定

乙型肝炎病毒(hepatitis B virus, HBV)是引起乙型肝炎的世界性流行病原菌。现场筛查HBV感染对预防和诊断HBV感染至关重要。本文开发了一种简单、直观、快速的 HBV-DNA 现场检测方法。该方法基于甜菜碱辅助重组酶聚合酶测定，然后通过侧向流测定 (BRPA-LF) 进行肉眼检测。结果表明，在重组酶聚合酶扩增（RPA）中，如果用未经纯化的血清样品进行检测，则容易发生非特异性扩增。此问题已通过向 RPA 反应添加 0.8 M 甜菜碱得到解决。结果表明，BRPA-LF 可以检测 50 μL 混合物中 1,000 拷贝的 HBV-DNA，并且对血清样品检测实现了 90% 的灵敏度和 100% 的特异性。

更新日期：2020-05-10

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