A novel paper based loop mediated isothermal amplification and lateral flow assay (LAMP‐LFA) for point‐of‐care detection of buffalo tissue origin in diverse foods

环介导等温扩增 分子生物学 色谱法 材料科学 生物 DNA 化学 生物化学
作者
Jyoti Jawla,Rajiv Ranjan Kumar,S.K. Mendiratta,Ravi Kant Agarwal,Praveen Singh,Vikas Saxena,Sarita Kumari,Dhananjay Kumar
出处
期刊:Journal of Food Safety [Wiley]
卷期号:43 (3) 被引量:6
标识
DOI:10.1111/jfs.13038
摘要

Abstract In this study, we have designed a paper based loop‐mediated isothermal amplification‐lateral flow (LAMP‐LF) Assay that integrated the use of self‐designed lyophilized paper LAMP buttons as ready‐made LAMP master‐mix for on‐site detection of tissue of buffalo origin. Based on mitochondrial cytochrome b gene sequences, a pair of LAMP primers and probes were selected and designed, respectively. Herein, we optimized LAMP reaction components and thermal protocol to evaluate LAMP amplification in various paper matrices frequently used in diagnostics (cellulose, glass fiber, and nitrocellulose). The hybridization conditions of probes, as well as other components of paper LAMP‐LF Assay, were further optimized. Our low‐cost paper based system demonstrated high specificity and was able to detect as low as 10 fg of target DNA. The efficiency and accuracy of the assay were determined targeting coded meat/tissue samples, binary meat admixture, processed meat samples, Phire Animal Tissue Direct PCR kit, and by investigating the consistency between paper based LAMP‐LFA and conventional PCR outcomes. Furthermore, the paper platform was allowed for refrigeration‐temperature storage to determine its ability for deployment in low settings areas. In the current investigation, the time required from sample collection to detection was approximately 2–3 hr.
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