Application of spectrophotometry to evaluate the concentration of purified White Spot Syndrome Virus.
White Spot Syndrome Virus (WSSV) is a highly virulent pathogen of shrimp. In previous work, a simple and efficient method has been established in our laboratory to purify intact WSSV virions from infected crayfish tissues. To perform studies of WSSV infection mechanism, pathogenesis and gene function by using this purified virion, quantitative assay for the virus becomes increasingly important. In this study, the optical density of the purified virion samples was measured at 600nm wavelength using spectrophotometer and the corresponding concentration was counted by transmission electron microscopy. The statistical results revealed a high correlation between optical density and concentration of WSSV virions (r=0.993; n=5). Finally, a conversion coefficient "f" (3.34x10(8)virions/microl) was obtained and a formula was established: C (virions/microl)=fOD(600)=3.34x10(8)xOD(600), which can be conveniently used to convert the optical density of purified WSSV preparation into the virion concentration.
Zhou
Qing
Q
State Key Laboratory of Virology, Section of Molecular Virology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China.
Qi
Yi-Peng
YP
Yang
Feng
F
D000818:Animals; D003400:Astacoidea / Q000821:virology*; D046529:Microscopy, Electron, Transmission; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods*; D046848:White spot syndrome virus 1 / Q000302:isolation & purification* / Q000648:ultrastructure