tetano
Editor, Senior Moderator
Methods Mol Biol. 2014;1200:107-20. doi: 10.1007/978-1-4939-1292-6_10.
A simple viral neuraminidase-based detection for high-throughput screening of viral hemagglutinin-host receptor specificity.
Sriwilaijaroen N1, Suzuki Y.
Author information
Abstract
The correlation between precise interactions of influenza A virus hemagglutinins with host cell surface glycans having terminal sialic acids and host range specificity has provoked the development of a high-throughput viral-receptor specificity assay. Here, we describe the use of the virus itself as a specific antibody coupled to enzymes (virus with neuraminidase spikes) for determining its binding specificity to glycans, a strategy that reduces not only the cost but also the tedious steps of adding primary and secondary antibodies and washing between each step. All of the steps, including coating the glycopolymers onto microtiter plates, virus binding, and visual and quantitative detection of fluorescence products that correlate well with the amount of glycan-bound viruses, can be done within 3 h. This simple, rapid, sensitive, and reliable strategy is an ideal method for detection of high-throughput influenza virus receptor-binding preference not only for studies on viral evolution and transmission but also for viral surveillance in pandemic preparedness, leading to efficient prevention and control of the disease.
PMID:
25117229
[PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/25117229
A simple viral neuraminidase-based detection for high-throughput screening of viral hemagglutinin-host receptor specificity.
Sriwilaijaroen N1, Suzuki Y.
Author information
Abstract
The correlation between precise interactions of influenza A virus hemagglutinins with host cell surface glycans having terminal sialic acids and host range specificity has provoked the development of a high-throughput viral-receptor specificity assay. Here, we describe the use of the virus itself as a specific antibody coupled to enzymes (virus with neuraminidase spikes) for determining its binding specificity to glycans, a strategy that reduces not only the cost but also the tedious steps of adding primary and secondary antibodies and washing between each step. All of the steps, including coating the glycopolymers onto microtiter plates, virus binding, and visual and quantitative detection of fluorescence products that correlate well with the amount of glycan-bound viruses, can be done within 3 h. This simple, rapid, sensitive, and reliable strategy is an ideal method for detection of high-throughput influenza virus receptor-binding preference not only for studies on viral evolution and transmission but also for viral surveillance in pandemic preparedness, leading to efficient prevention and control of the disease.
PMID:
25117229
[PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/25117229