Introduction: Epidemic Japanese encephalitis is one of the most important zoonotic diseases that cause central nervous system damage. The vaccination has become the most effective and economical measure for its control. Hence, real-time monitoring of Japanese encephalitis virus (JEV) proliferation is crucial to optimize virus inoculation, culturing conditions, and virus harvest time. Methods: The proliferation dynamics of JEV in BHK-21 cells was studied by combining the established quantitative PCR method with the conventional TCID₅₀ assay in this study. Results: The proliferation curve determined by the 2 methods has a definite parallel relationship, but the quantitative real-time PCR method (4 h) is faster and more sensitive than the TCID₅₀ method (3–4 days). The determination results of TCID₅₀ showed that the highest viral titer was 10^5.44 TCID₅₀/0.1 mL and 10^4.86 TCID₅₀/0.1 mL in cell suspension and culture supernate, respectively, while the virus RNA copies reached the peak at 1.0 × 10^7.5 copies/µL and 1.0 × 10^5.6 copies/µL in cell suspension and culture supernate, respectively. Conclusion: The comprehensive analysis showed that the best time for JEV proliferation in BHK-21 cell was 60 h post infection.
Intervirology

中文翻译：

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JEV在BHK-21细胞中动态增殖的研究


简介：流行性日本脑炎是引起中枢神经系统损害的最重要的人畜共患疾病之一。疫苗接种已成为控制其最有效、最经济的措施。因此，实时监测日本脑炎病毒（JEV）增殖对于优化病毒接种、培养条件和病毒收获时间至关重要。方法：本研究将建立的定量PCR方法与常规TCID ₅₀测定相结合，研究JEV在BHK-21细胞中的增殖动态。结果： 2种方法测定的增殖曲线具有明确的平行关系，但实时定量PCR法（4 h）比TCID ₅₀法（3~4 d）更快、更灵敏。 TCID ₅₀测定结果显示，细胞悬液和培养上清液中病毒滴度最高分别为10 ^5.44 TCID ₅₀ /0.1 mL和10 ^4.86 TCID ₅₀ /0.1 mL，病毒RNA拷贝数达到峰值为1.0 × 10 ^7.5细胞悬液和培养上清液中分别为 1.0 × 10 5.6 拷贝/μL 和 1.0 × 10 ^5.6拷贝/μL。结论：综合分析表明，JEV在BHK-21细胞中增殖的最佳时间为感染后60 h。
 病毒间学