Zhang Bo, Zheng Lan, Huang Yu-Wen, Mo Qin, Wang Xun, Qian Kai-Cheng
Shanghai Blood Center Graduate Student Training Base, East China Normal University, Shanghai 200051, China.
Bing Du Xue Bao. 2009 Jul;25(4):286-90.
To investigate the feasibility of using Real-Time PCR to evaluate the effectiveness of Sindbis virus inactivation by Methylene Blue with visible light. Sindbis virus was treated by Methylene Blue with different intensity of visible light and the transcribed cDNA was quantified by Real-Time PCR. Residual infectivity of treated virus was tested by cell infection method as parallel control at the same time. The residual infectivity of virus decreased from 6.50 lgTCID50/mL to under the limit of detection as light intensity increased. Meanwhile, the quantity of virus cDNA decreased significantly (P < 0.05), which correlated to the decline of virus infectivity (R2 > 0.98). Methylene Blue with visible light could cause lesion to nucleic acid of Sindbis virus, the extent of which was light intensity-dependent and correlated to the decrease of virus infectivity. The results demonstrated that Real-Time PCR can be a useful tool for evaluating effect of virus inactivation after Methylene Blue treatment with light.
为研究使用实时荧光定量聚合酶链反应（Real-Time PCR）评估亚甲蓝联合可见光对辛德毕斯病毒灭活效果的可行性。用不同强度可见光处理亚甲蓝与辛德毕斯病毒，通过实时荧光定量聚合酶链反应对转录的互补脱氧核糖核酸（cDNA）进行定量。同时，采用细胞感染法检测处理后病毒的残余感染性作为平行对照。随着光强度增加，病毒的残余感染性从6.50 lgTCID50/mL降至检测限以下。与此同时，病毒cDNA数量显著减少（P < 0.05），这与病毒感染性的下降相关（决定系数R2 > 0.98）。亚甲蓝联合可见光可导致辛德毕斯病毒核酸损伤，损伤程度依赖于光强度且与病毒感染性降低相关。结果表明，实时荧光定量聚合酶链反应可作为评估亚甲蓝光照处理后病毒灭活效果的有用工具。
