免疫捕获联合基质辅助激光解吸电离飞行时间质谱快速检测 1 型登革病毒。

Immunocapture couples with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid detection of type 1 dengue virus.

机构信息

Department of Biological Science and Technology, National Chiao Tung University, Hsin-Chu, Taiwan.

出版信息

J Chromatogr A. 2013 May 3;1288:21-7. doi: 10.1016/j.chroma.2013.02.030. Epub 2013 Feb 28.

DOI:10.1016/j.chroma.2013.02.030

Abstract

A facile method for accurate detection of type 1 dengue virus (DV1) infection from complex biological mixtures, using type specific immunocapture coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), was developed. The biological mixtures were treated with magnetic particles coated with a monoclonal antibody directly against type 1 dengue virus. After immunocapture purification, the DV1 was eluted with 30% acetic acid, directly spotted with seed-layer method, and analyzed by MALDI-TOF MS for DV1 capsid protein. The detection limit of the assay was ∼10(5)pfu/mL by MALDI-TOF MS. The immunocapture could unambiguously differentiate the DV1 from other serotypes of the dengue viruses and Japanese encephalitis virus, and could be used as a specific probe to detect DV1 from complex biological mixtures.

摘要

建立了一种简便的方法，可使用针对 1 型登革热病毒（DV1）的单克隆抗体包被的磁性颗粒从复杂的生物混合物中准确检测 1 型登革热病毒（DV1）感染，该方法结合了免疫捕获和基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）。将生物混合物用直接针对 1 型登革热病毒的单克隆抗体包被的磁性颗粒进行处理。免疫捕获纯化后，用 30%乙酸洗脱 DV1，用种晶层法直接点样，用 MALDI-TOF MS 分析 DV1 衣壳蛋白。该方法的检测限为 MALDI-TOF MS 约 10(5)pfu/mL。免疫捕获可明确区分 DV1 与其他登革热病毒和日本脑炎病毒的血清型，可作为从复杂生物混合物中检测 DV1 的特异性探针。