Study on high-throughput respiratory tract virus detection method based SPR / 国际检验医学杂志

ABSTRACT

Objective To develop a biosensor method with strong specificity and high‐throughput by combining with the surface plasmon resonance (SPR) and gene chip technique and aiming at 9 kinds of common respiratory tract viruses including influenza A and influenza B ,(Influ A ,B) ,H1N1 ,respiratory syncytial virus (RSV) ,parainfluenza virus 1 -3 (PIV1 -3) ,adenovirus (ADV) and coronavirus (SARS) leading to severe acute respiratory syndrome .Methods Firstly the software primer 5 was used to design the specific primer and probe of related viruses in the conserved sequence ;the designed nine kinds of corresponding respiratory virus probes were immobilized in the specific region of SPR chip after chemical modification .The SPR technique was applied to conduct the real time monitoring the hybridization process of the probe with the PCR products .Finally the signal amplification was realized by the biotin and streptavidin system .Results The designed gene chip could detect 9 kinds of respiratory tract viruses by high‐throughput with better detection specificity ;the chip surface could be reutilized after certain regeneration condition ,which avoided the influence of intra‐batch difference on the results ;the detection sensitivity reached the nanomole level .Conclusion The prelimi‐nary study results demonstrate that using the SPR biosensor technique to establish a high‐‐throughput detection of respiratory tract viruses has some practicability and feasibility ,and is expected to become a rapid ,large scale and high‐ throughput measure for screening respiratory tract viruses with good application prospect .