The Application Of Microfluidic Chip Technology In Brucella Detection

Objective To establish and evaluate the method of detecting Brucella nucleic acid based on microfluidic chip(MC)technology,so as to improve the diagnostic ability of Brucella.Method To design a fully automatic technical process for the detection of Brucella DNA by the combination of MC nucleic acid extraction,PCR and DNA reverse dot blot hybridization.Design the compliance test between microfluidic automatic nucleic acid detection(MC-PCR)and gene sequencing,and compare it with real-time fluorescence quantitative nucleic acid amplification detection(q PCR)and Taq Man probe detection;To evaluate the repeatability,accuracy,specificity and minimum detection limit of Brucella detected by MC-PCR.Results The extraction efficiency of nucleic acid by MC method was the second,and there was significant difference compared with the six methods in the control group(P <0.05).The kappa value of MC-PCR and gene sequencing was 0.659,and the detection was highly consistent;The accuracy rate was 100%;The negative rate of specific results was100%;Brucella could be detected in the repeatability results;When the detection limit was825 copies/ml,there was significant difference in the detection rate between MC-PCR,Taq Man and q PCR(P < 0.05).Conclusion The detection method of Brucella nucleic acid by MC-PCR is safe,fast,timely and accurate,and can be used in clinic.This will improve the diagnosis rate of brucellosis and play an important role in the prevention and treatment of brucellosis.