tailieunhanh - báo cáo hóa học:" Rapid label-free identification of mixed bacterial infections by surface plasmon resonance"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí sinh học đề tài : Rapid label-free identification of mixed bacterial infections by surface plasmon resonance | Wang et al. Journal of Translational Medicine 2011 9 85 http content 9 1 85 JOURNAL OF TRANSLATIONAL MEDICINE RESEARCH Open Access Rapid label-free identification of mixed bacterial infections by surface plasmon resonance 1 1 1 2 1 2 IA z- 1 1 Jue Wang Yang Luo Bo Zhang Ming Chen Junfu Huang Kejun Zhang Weiyin Gao Weiling Fu Tianlun Jiang3 and Pu Liao4 Abstract Background Early detection of mixed aerobic-anaerobic infection has been a challenge in clinical practice due to the phenotypic changes in complex environments. Surface plasmon resonance SPR biosensor is widely used to detect DNA-DNA interaction and offers a sensitive and label-free approach in DNA research. Methods In this study we developed a single-stranded DNA ssDNA amplification technique and modified the traditional SPR detection system for rapid and simultaneous detection of mixed infections of four pathogenic microorganisms Pseudomonas aeruginosa Staphylococcus aureus Clostridium tetani and Clostridium perfringens . Results We constructed the circulation detection well to increase the sensitivity and the tandem probe arrays to reduce the non-specific hybridization. The use of 16S rDNA universal primers ensured the amplification of four target nucleic acid sequences simultaneously and further electrophoresis and sequencing confirmed the high efficiency of this amplification method. No significant signals were detected during the single-base mismatch or non-specific probe hybridization P . The calibration curves of amplification products of four bacteria had good linearity from nM to 100 nM with all R2 values of . The lowest detection limits were nM for P. aeruginosa nM for S. aureus nM for C. tetani and nM for C. perfringens. The SPR biosensor had the same detection rate as the traditional culture method P . In addition the quantification of PCR products can be completed within 15 min and excellent regeneration greatly reduces the .