Celltiter-Glo ATP荧光活性检测在登革病毒1型中和试验中的应用

Application of Celltiter-Glo ATP fluorescence analysis in type 1 Dengue virus neutralization test

目的利用Celltiter-Glo ATP荧光活性检测方法(CTG法)在登革病毒1型(DENV1)中和试验中的实验研究,探索出新的组织培养中和试验结果判断方法。方法 DENV1病毒与抗DENV1多克隆抗体进行中和试验感染Vero细胞,选择CTG法和传统的显微镜观察法判断CPE的程度,比较两种方法对细胞半数致死量(TCID50)和中和指数的判定,确定CTG法的准确性。结果显微镜观测TCID50=5.5,CTG法TCID50=5.31;显微镜观测、CTG检测法均为10-2.1为中和终点,两种方法获得的中和指数较一致。结论 CTG法可作为DENV1细胞组织培养中和试验的结果判断,并能减少实验结果评定中的人为误差。

OBjective To study new result judgment method in tissue culture neutralization test by applying Cel｜titer-Glo ATP fluorescence analysis（CTG） in type 1 dengue virus （DENV1） neutralization test. Methods DENV1 and anti DENV1 polyclonal antibody were applied to Veto cells for neutralization test, CTG method and traditional microscope observation method were used to determine degree of CPE. Half lethal dose （TCID50） and cell neutralization index by the above 2 methods were determined and compared to evaluate the accuracy of CTG method. Results TCID50 was 5.5 for traditional microscope observation method and 5.31 for CTG method. Neutralization titer of both methods was 10 ^-2.l. Neutralization titer was con- sistent for both methods. Conclusion CTG method can reduce human error in result judgment, which can be used as result judgment method in DENV1 tissue culture neutralization test.