Abstract: In order to improve the sensitivity of RT-qPCR method for detection of Newcastle Disease Virus （NDV）, the concentration of lysis buffer （guanidinium isothiocyanate） and washing solution 1 （guanidine hydrochloride） used in column extraction method for detection of NDV nucleic acid was optimized; the lysis effect of Triton-100 and EDTA on NDV particles was evaluated; the performance of the optimized column extraction method and the traditional Trizo] method in NDV nucleic acid extraction was compared by RT-qPCR. The results showed that the optimum concentration of lysis buffer （guanidinium isothiocyanate） was determined as 5 mol/L and that of solution 1 （guanidine hydrochloride） was determined as 5.5 mol/L; addition of 20 mmol/L EDTA in lysis buffer exhibited an inhibitory effect on the extraction of NDV nucleic acid; addition of 0.8% of Triton-100 in lysis buffer improved the extraction efficiency of NDV nucleic acid. Compare to the traditional Trizol method, the optimized column extraction method had higher extraction efficiency. The extraction method of NDV nucleic acid was optimized by this study.