Application of different nucleic acid rapid extraction method in the detection of Brucella abortus evaluated
Objective To compare the effects of different nucleic acid extraction methods on the detection rate of Brucella,and to provide technical reference for the rapid extraction and detection of Brucella abortus nucleic acid.Methods Magnetic bead method,direct cracking method and high temperature cracking method were used to extract nucleic acid from 20 whole blood samples and 10 joint cavity and serosal cavity effusion samples of Brucella infected patients in our hospital,and then real-time fluorescence RT-PCR was performed to evaluate the DNA extraction efficiency of this rapid nucleic acid extraction reagent.Results The positive detection rate of magnetic bead method was 100%,the median Ct value was 29.17(21.19～31.33),and the maximum intra-batch coefficient of varia-tion was 1.33%.The positive detection rate of direct cracking method was 63.33%,the median Ct value was 33.29(28.69～34.54),and the maximum coefficient of variation was 1.28%.The positive rate of high temperature cracking method was 100%,the median Ct value was 30.26(26.63～33.85),and the coefficient of variation in batch was the highest 1.21%.There was no significant difference in the detection rate between high temperature cracking method and magnetic bead method(P>0.05),but the detection rate of direct cracking method was significantly lower than that of magnetic bead method and high temperature cracking method(P<0.05).Conclusion The nucleic acid rapid extraction reagent can ensure the efficiency and repeatability of nucleic acid extraction under the condition of high temperature cracking,and has the advantages of simple operation,shorten detection time,and reduce laboratory biosafe-ty,so it can be applied to Brucella nucleic acid detection.
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