Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Use of a highly sensitive strand-specific quantitative PCR to identify abortive replication in the mouse model of respiratory syncytial virus disease | Bannister et al. Virology Journal 2010 7 250 http content 7 1 250 VIROLOGY JOURNAL RESEARCH Open Access Use of a highly sensitive strand-specific quantitative PCR to identify abortive replication in the mouse model of respiratory syncytial virus disease Richard Bannister Deborah Rodrigues Edward J Murray Carl Laxton Mike Westby Helen Bright Abstract Background The BALB c mouse is commonly used to study RSV infection and disease. However despite the many advantages of this well-characterised model the inoculum is large viral replication is restricted and only a very small amount of virus can be recovered from infected animals. A key question in this model is the fate of the administered virus. Is replication really being measured or is the model measuring the survival of the virus over time To answer these questions we developed a highly sensitive strand-specific quantitative PCR QPCR able to accurately quantify the amount of RSV replication in the BALB c mouse lung allowing characterisation of RSV negative and positive strand RNA dynamics. Results In the mouse lung no increase in RSV genome was seen above the background of the original inoculum whilst only a limited transient increase 1 log in positive strand replicative intermediate RI RNA occurred. This RNA did however persist at detectable levels for 59 days post infection. As expected ribavirin therapy reduced levels of infectious virus and RI RNA in the mouse lung. However whilst Palivizumab therapy was also able to reduce levels of infectious virus it failed to prevent production of intracellular RI RNA. A comparison of RSV RNA kinetics in human A549 and mouse KLN205 cell lines demonstrated that RSV replication was also severely delayed and impaired in vitro in the mouse cells. Conclusions This is the first time that such a sensitive strand-specific QPCR technique has been to the RSV mouse system. We have accurately quantified the restricted and abortive nature of RSV replication in the .
