Detection of influenza A virus RNA in birds by optimized Real-Time PCR system

Objective:To evaluate the use of Real-Time PCR system based on specific amplification of matrix protein gene fragment for influenza A virus RNA detection in cloacal swabs from wild birds.Methods:Sensitivity,specificity and reproducibility of analysis results were identified. Study of cloacal swabs from wild birds for influenza A virus presence was performed.Results: Reproducibility of low concentrations of virus detection in samples by Real-Time PCR was significantly higher than that of detection based on cytopathic effect of viruses grown on MDCK cell culture.Conclusions:Real-Time PCR system for influenza A virus RNA detection is developed and applied for virus surveillance study.

3-乙酰基-11-羰基-β-乙酰乳香酸在Caco-2和MDCK细胞模型中的吸收研究

目的研究乳香提取物中3-乙酰基-11-羰基-β-乙酰乳香酸(AKBA)在Caco-2、MDCK-MDR1和MDCK-Wild细胞模型中的吸收转运机制。方法利用Caco-2、MDCK-MDR1和MDCK-Wild细胞模型,研究AKBA由细胞层顶端(AP)→基底端(BL)和BL→AP的双向转运过程;采用LC-MS/MS法测定AKBA的量,计算表观渗透系数(Papp)。结果在Caco-2细胞模型中,50μmol/L AKBA由AP→BL、BL→AP的Papp分别为7.9×10 7、1.5×10 7cm/s,在MDCK-MDR1细胞模型中,50μmol/LAKBA由AP→BL、BL→AP的Papp分别为2.6×10 7、0.8×10 7cm/s,在MDCK-Wild细胞模型中,50μmol/LAKBA由AP→BL、BL→AP的Papp分别为2.4×10 7、0.6×10 7cm/s,3种细胞模型中外排率均小于2。结论 AKBA在肠道中吸收不良,不是P-糖蛋白的底物,推测其通过摄入型主动转运和被动扩散两种机制透过小肠上皮细胞。