A simple method for Alexa Fluor dye labelling of dengue virus.
Dengue virus causes frequent and cyclical epidemics throughout the tropics, resulting in significant morbidity and mortality rates. There is neither a specific antiviral treatment nor a vaccine to prevent epidemic transmission. The lack of a detailed understanding of the pathogenesis of the disease complicates these efforts. The development of methods to probe the interaction between the virus and host cells would thus be useful. Direct fluorescence labelling of virus would facilitate the visualization of the early events in virus-cell interaction. This report describes a simple method of labelling of dengue virus with Alexa Fluor succinimidyl ester dye dissolved directly in the sodium bicarbonate buffer that yielded highly viable virus after labelling. Alexa Fluor dyes have superior photostability and are less pH-sensitive than the common dyes, such as fluorescein and rhodamine, making them ideal for studies on cellular uptake and endosomal transport of the virus. The conjugation of Alexa Fluor dye did not affect the recognition of labelled dengue virus by virus-specific antibody and its putative receptors in host cells. This method could have useful applications in virological studies.
Zhang
Summer Li-Xin
SL
Defence Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, 117510 Singapore, Singapore.
Tan
Hwee-Cheng
HC
Hanson
Brendon J
BJ
Ooi
Eng Eong
EE
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003716:Dengue Virus / Q000302:isolation & purification* / Q000502:physiology; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D008027:Light; D050296:Microbial Viability / Q000187:drug effects; D009930:Organic Chemicals; D013194:Staining and Labeling / Q000379:methods*; D014773:Virology / Q000379:methods*
C417664:Alexa594; D005456:Fluorescent Dyes; D009930:Organic Chemicals