CAJ | 학술논문

目的：对5种DNA提取方法进行比对，为选择适用于痰中结核分枝杆菌DNA的提取方法提供依据。方法：采用PureLink genomic DNA kits (PLKm)、苯酚与氯仿法(PCm)、Chelex-100法(Cm)、磷酸盐缓冲液法(PBm)及硅珠法(SBm)5种方法提取痰标本中的DNA，对提取产物的浓度、纯度等技术指标进行分析，并对核酸扩增结果进行统计。结果：DNA浓度均值由高到低依次是PBm、Cm、PCm、SBm、PLKm，组间比较差异有统计学意义，两两比较发现PLKm与PCm、Cm、PBm之间差异有统计学意义，但Cm与SBm间的比较有统计学意义；5种提取方法的OD260：OD280比值均值与1.8的接近程度依次是PLKm、PCm、Cm、PBm、SBm，组间比较差异有统计学意义，两两比较发现除SBm与另外4种方法间存的差异具有统计学意义外，其余各组间的比较均无统计学意义；采用PLKm、PCm及Cm提取的产物均扩增出阳性结果，而PBm、SBm提取产物分别有18例和11例未获得阳性结果，经Mc-Nemar x2检验方法间的差异有统计学意义。结论：Chelex-100提取痰标本中结核分枝杆菌DNA是一种简便、省时、廉价适用于基层结核病实验室进行核酸检测的方法。
목적：대5충DNA제취방법진행비대，위선택괄용우담중결핵분지간균DNA적제취방법제공의거。방법：채용PureLink genomic DNA kits (PLKm)、분분여록방법(PCm)、Chelex-100법(Cm)、린산염완충액법(PBm)급규주법(SBm)5충방법제취담표본중적DNA，대제취산물적농도、순도등기술지표진행분석，병대핵산확증결과진행통계。결과：DNA농도균치유고도저의차시PBm、Cm、PCm、SBm、PLKm，조간비교차이유통계학의의，량량비교발현PLKm여PCm、Cm、PBm지간차이유통계학의의，단Cm여SBm간적비교유통계학의의；5충제취방법적OD260：OD280비치균치여1.8적접근정도의차시PLKm、PCm、Cm、PBm、SBm，조간비교차이유통계학의의，량량비교발현제SBm여령외4충방법간존적차이구유통계학의의외，기여각조간적비교균무통계학의의；채용PLKm、PCm급Cm제취적산물균확증출양성결과，이PBm、SBm제취산물분별유18례화11례미획득양성결과，경Mc-Nemar x2검험방법간적차이유통계학의의。결론：Chelex-100제취담표본중결핵분지간균DNA시일충간편、성시、렴개괄용우기층결핵병실험실진행핵산검측적방법。
Objective Comparison five DNA extraction methods of sputum specimen, To select the applicable DNA extraction method of Mycobacterium tuberculosis in basic tuberculosis research laboratories. Methods Using the five methods to extract DNA of Mycobacterium tuberculosis from sputum, PureLink genomic DNA kits (PLKm), phenol and chloroform method (PCm), Chelex-100 method (Cm), Phosphate buffer method (PBm) and silica beads method (SBm), to analyze concentration, purity and other technical indicators of the extraction product, and to do the statistical analysis of nucleic acid amplification results. Results The Average DNA concentration values from high to low in turn is PBm, Cm, PCm, SBm and PLKm, there was statistically significant difference between the groups.The Average values of OD260:OD280 Proximity to 1.8 from high to low in turn is PLKm, PCm, Cm, PBm and SBm, there was statistically significant dif-ference between the groups(P=0.001),there were statistically significant difference between SBm and the other methods (P<0.05) but conversely on other groups by comparison between any two means (P>0.05);The extraction products amplified of PLKm, PCm and Cm were positive, while 18 and 11 samples without obtaining positive results by the PBm and SBm , have statistical significance difference between the groups. Conclusion Using Chelex-100 to extract DNA of Mycobacterium tuberculosis from sputum is a simple, time-saving and cheap method for nucleic acid detection in basic tuberculosis re-search laboratories.