3种核酸提取方法及3种实时荧光定量PCR仪检测结果的比较

Comparative study on three methods of nucleic acid extraction and three kinds of real-time fluorescence quantitative PCR instrument

目的 探讨不同核酸提取方法以及不同实时荧光定量PCR仪检测结果之间的差异.方法 随机挑选呼吸道、肠道病毒阳性标本各25份,采用3种方法（方法A、B、C）进行核酸提取,比较不同核酸提取方法之间的差异;随机挑选呼吸道、肠道病毒阳性核酸各25份,在3种实时荧光定量PCR仪（仪器A、B、C）上进行实时荧光定量PCR,比较不同实时荧光定量PCR仪之间的差异.检测结果采用定量资料的随机区组方法分析.结果 3种核酸提取方法检测结果存在差异（x2=42.9162,P〈0.001）,其中方法A、B,方法B、C之间的差异具有统计学意义（Z=7.025,P〈0.001;Z=7.9,P〈0.001）,方法A、C之间的差异无统计学意义（Z=0.837,P=0.3816〉0.05）.3种实时荧光定量PCR仪的检测结果也存在差异（x2=23.773,P〈0.001）,其中仪器A、B,仪器A、C之间的差异有统计学意义（Z=5.70,P〈0.001;Z=6.45,P〈0.001）,仪器B、C之间的差异无统计学意义（Z=0.75,P=0.4533〉0.05）.结论 在相同条件下,用不同的核酸提取方法,以及用不同的实时荧光定量PCR仪检测的结果均有差异,在实际工作中要综合考虑评价检测结果.

Objective To explore the differences among three methods of nucleic acid extraction and three kinds of real-time fluorescence quantitative PCR instrument.Methods Twenty-five respiratory virus nucleic acid and 25 enterovirus nucleic acid positive samples were with selected at random and nucleic acids were extracted by using three methods （method A, B, and C）.The results among different methods were analyzed by randomized block design.25 respiratory viral nucleic acid positive specimens and enterovirus nucleic acid positive samples were detected by using three kinds of real-time fluorescence quantitative PCR instrument （instrument A, B, and C）.The results among different instruments were analyzed by randomized block design.Results There was a significant difference among three methods of nucleic acid extraction in results（x2=42.9162, P〈0.001）, in which method A and C had not significant difference（Z=0.837,P=0.3816〉0.05）, while method A vs.B, B vs.C were significantly different（Z=7.025,P〈0.001;Z=7.9,P〈0.001）.There was also a significant difference among three kinds of real-time fluorescence quantitative PCR instrument in results（x2=23.773, P〈0.001）, in which instrument B and C had no significant difference（Z=0.75,P=0.4533〉0.05）, while instrument A vs.B, A vs.C were significantly different（Z=5.70,P〈0.001;Z=6.45,P〈0.001）.Conclusions There is difference among different methods and instruments in the test results under the same condition, which call for options in practical work according to need.