Analysis of different nucleic acid extraction methods on the verification of HBV-DNA detection performance
Objective To evaluate the performance of two hepatitis B virus(HBV)-DNA extraction methods and two detection reagents,and to help select and optimize the extraction reagents and detection rea-gents.Methods In April 2023,Da'an automatic nucleic acid extractor extraction method(magnetic bead meth-od)and manual extraction method(one-step method)were used to detect HBV-DNA with Da'an and Shengx-iang reagents.The precision,accuracy,linear range,detection limit and anti-interference ability were verified and evaluated.Results The results showed that the detection results of Da'an automatic nucleic acid extrac-tion instrument extraction of Da'an reagent detection,manual extraction of Da'an reagent detection and manu-al extraction of Shengxiang reagent detection were up to the standard in terms of precision,accuracy,linear range and detection limit.Shengxiang reagent was extracted by Da'an automatic nucleic acid extraction instru-ment,coefficient of variability＞5％in the low value detection,the minimum detection limit verification was not qualified.In terms of anti-interference ability,the samples with 2.0 g/dL hemoglobin concentration ex-tracted by the automatic nucleic acid extraction instrument were not affected by the detection results of Da'an and Shengxiang reagents.Samples with triglyceride concentration up to 3 000 mg/dL were extracted manual-ly,and the detection results of both reagents were not affected.Conclusion Mixing of extraction and detection reagents from different manufacturers should be avoided.The results of quantitative detection of HBV-DNA by the integrated reagents of Da'an and Shengxiang met the requirements.
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