Abstract

The current study made an attempt to develop a novel surface plasmon resonance (SPR) sensor using wheat germ agglutinin as a biomolecule for the rapid detection of Listeria monocytogenes. A selective biomolecular interaction of wheat germ agglutinin (WGA) lectin with surface carbohydrate components of L. monocytogenes was used as a biorecognition principle. From the previous studies based on agglutination and sugar inhibition principle, we have selected WGA as a highly selective ligand for L. monocytogenes-WGA biomolecular interaction study using a SPR (Biacore 3000) sensor. Under optimized assay conditions, L. monocytogenes strains have generated maximum response of 479 ± 49 resonance unit (RU) at 7.4 log cfu/100 μl with a least cross-reactivity in presence of both G+ve and G−ve Bacterial contaminants. The limit of detection of 3.25 log CFU/100 μl was obtained for the detection of L. monocytogenes by lectin immobilized WGA CM5 chip surface using SPR. Further, the WGA-immobilized chip was evaluated with milk samples enriched in Listeria selective enrichment medium and the response (RU) was again found to be significant up to 3.0 log CFU/100 μl.

中文翻译：

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小麦胚芽凝集素通过表面等离子体共振快速检测牛奶中的李斯特菌

摘要

当前的研究试图开发一种新型表面等离振子共振（SPR）传感器，该传感器使用小麦胚芽凝集素作为生物分子来快速检测单核细胞增生李斯特菌。小麦胚芽凝集素（WGA）凝集素与单核细胞增生李斯特氏菌表面碳水化合物成分的选择性生物分子相互作用被用作生物识别原理。从以前的基于凝集和糖抑制原理的研究中，我们选择WGA作为单核细胞增生李斯特氏菌的高选择性配体-使用SPR（Biacore 3000）传感器进行WGA生物分子相互作用的研究。在优化的测定条件下，单核细胞增生李斯特菌在G + ve和G-ve细菌污染物同时存在时，菌株在7.4 log cfu / 100μl时产生的最大响应为479±49共振单位（RU），交叉反应最少。通过使用SPR通过凝集素固定的WGA CM5芯片表面检测单核细胞增生李斯特氏菌，获得了3.25 log CFU / 100μl的检测限。此外，用富含利斯特氏菌选择性富集培养基的牛奶样品评估了WGA固定化芯片，并且再次发现响应（RU）高达3.0 log CFU / 100μl。