Quantitative detection of the loop mediated isothermal amplification gene in the loopmediated isothermal amplification of Japanese encephalitis and dengue fever / 国际检验医学杂志

Objective To establish a loop-mediated isothermal amplification(LAMP) quantitative method for rapid detection of Japanese encephalitis and dengue fever virus.Methods According to the LAMP principle,design primers for LAMP detection and reaction system,establish LAMP detection method,and to evaluate the linear relationship between initial copy number and the specificity,sensitivity,repeatability and the reaction time(fluorescence signal value of 1×104 corresponding time).Results One sets of LAMP primers could be used to complete the detection work in 0.5 h.The sensitivity of LAMP detection technology was 10 times higher than that of the classical PCR technology,and no cross reaction with other viruses,and the coefficient of variation of the average test was less than 5%.There was a good linear relationship between cycle threshold and template concentration.Conclusion This method has high specificity,sensitivity,simple operation,which is easy to get the results,low equipment requirements and rapid,suitable for primary health institutions and the field inspection agencies for wide applications.

Research and development of genetic diagnostic method of Staphyloccocus aureus based on loop-mediated isothermal amplification / 中华微生物学和免疫学杂志

Objective To develop a method of loop-mediated isothermal amplification(LAMP) to Staphyloccocus aureus rapidly, specifically, sensitively and simply suited for the primary health agency. Methods According to conserved nucleotide of Staphyloccocus aureus and principle of LAMP, we designed a set of LAMP primers and set up an LAMP reaction system. We evaluated the specificity, sensitivity and re-peatability of the method. In addition,we evaluated the linearity between initial template copies 1g value and reaction time (the time when the fluorescent value is 1×10~4). Results The optimal assay showed that it was no cross-reaction with other closely related members of pathogens, and was 10 times more sensitive than PCR. The coefficient of variance between tests was less than 5% ,and the kinetics curves showed a good line-arity between initial template copies lg value and reaction time(r~2=0. 9501). The detection activity could be finished within 1 h with the sensitivity of LAMP was 100% and the specificity was 94.4%, and the accuracy was 96.6%. Conclusion These findings demonstrated that the LAMP had the potential clinical application for detection and differentiation of Staphyloccocus aureus in the public health agency for its sensitive, specific and simple feature.