A new method for quantifying white spot syndrome virus: Experimental challenge dose using TaqMan real-time PCR assay.
White spot syndrome virus (WSSV) is an important pathogen in shrimp aquaculture. The susceptibility of crayfish (Procambarus clarkii) was assessed by means of serial dilutions of a solution containing WSSV. A TaqMan real-time PCR was used to quantify the WSSV challenge dose in P. clarkii. The results showed that WSSV copies could be detected at concentrations from 1.365×10(4) to 1.129×10(9) copies/μl. The viral infectivity (LD(50)), measured as the mortality of infected crayfish, indicated 60% mortality in the 10(5) dilution group (1.524×10(5) copies/μl). TaqMan real-time PCR represents a novel standard method, based on the by quantitation of WSSV copies, for determining the appropriate concentration of WSSV for use in infection experiments.
College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Lin'an 311300, China. zhufei1998@yahoo.cn
Fei
F
Zhu
Haizhi
H
Quan
D000818:Animals; D003400:Astacoidea / Q000821:virology*; D007928:Lethal Dose 50; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D016019:Survival Analysis; D019562:Viral Load / Q000379:methods*; D046848:White spot syndrome virus 1 / Q000302:isolation & purification* / Q000472:pathogenicity