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  • 标题:Investigation of the “Antigen Hook Effect” in Lateral Flow Sandwich Immunoassay: The Case of Lumpy Skin Disease Virus Detection
  • 本地全文:下载
  • 作者:Simone Cavalera ; Giulia Pezzoni ; Santina Grazioli
  • 期刊名称:Biosensors
  • 电子版ISSN:2079-6374
  • 出版年度:2022
  • 卷号:12
  • 期号:9
  • DOI:10.3390/bios12090739
  • 语种:English
  • 出版社:MDPI Publishing
  • 摘要:Lumpy skin disease (LSD) is an infectious disease affecting bovine with severe symptomatology. The implementation of effective control strategies to prevent infection outbreak requires rapid diagnostic tools. Two monoclonal antibodies (mAbs), targeting different epitopes of the LSDV structural protein p32, and gold nanoparticles (AuNPs) were used to set up a colorimetric sandwich-type lateral flow immunoassay (LFIA). Combinations including one or two mAbs, used either as the capture or detection reagent, were explored to investigate the hook effect due to antigen saturation by the detector antibody. The mAb-AuNP preparations were optimized by a full-factorial design of experiment to achieve maximum sensitivity. Opposite optimal conditions were selected when one Mab was used for capture and detection instead of two mAbs; thus, two rational routes for developing a highly sensitive LFIA according to Mab availability were outlined. The optimal LFIA for LSDV showed a low limit of detection (10 3.4 TCID 50/mL), high inter- and intra-assay repeatability (CV% < 5.3%), and specificity (no cross-reaction towards 12 other viruses was observed), thus proving to be a good candidate as a useful tool for the point-of-need diagnosis of LSD.
  • 关键词:point-of-care test;rapid diagnosis;infectious diseases;single-epitope sandwich;double-epitope sandwich;lumpy skin disease
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