Objective In order to establish a ring-mediated isothermal amplification detection system for human H1N1 influenza virus,the nucleic acid amplification efficiency of H1N1 virus was used as an indicator,and to explore optimal reaction conditions for loop mediated isothermal amplification(LAMP)detection of H1N1 influenza virus by response surface analysis optimization.Methods Based on the principle of Box-Benhnken response design,the key factors affecting LAMP reaction (betaine concentration,primer concentration,and reaction temperature) were optimized by the three factors level response surface method.At the same time,60 clinical nucleic acid specimens of confirmed influenza A and 60 nucleic acid samples of healthy people were selected and evaluated by polymerase chain reaction (PCR) assay and LAMP assay comparatively (the evaluation indexes included sensitivity,specificity and accuracy).Results The response surface regression model was able to better predict the relationship between the LAMP detection effect and betaine concentration,primer concentration and reaction temperature,and the optimized LAMP improved the detection sensitivity and shortened the reaction time.Compared with PCR results,the LAMP assay achieved 100% sensitivity,98.9% specificity and 99.2%accuracy,and the lower limit of detection was up to 100 copies/ml.Conclusion The LAMP assay for influenza A (H1N1)established in the present study is a highly efficient,sensitive,and specific assay that does not require complex equipment,and it is suitable for the detection of H1N1 in basic community units or at home,which is useful for the monitoring of H1N1 and the post-outbreak epidemic.It is of great significance for the monitoring of influenza A disease and the emergency detection of outbreaks.