摘要
目的筛选解脲脲原体（ＵＵ）尿素酶基因上的不同引物及扩增模板区，使扩增敏感性达到最佳。方法以ＵＵ尿素酶基因为靶序列，设计５对引物，用聚合酶链反应（ＰＣＲ）扩增ＵＵ１～１４个血清型和系列稀释的ＵＵ８ＤＮＡ，用ＯＬＩＧＯ程序分析引物序列与ＰＣＲ敏感性间的关系。结果不同种的引物对１４个血清型的扩增结果有差异。引物Ｕ１＋Ｂ、Ｕ１＋２、ＵＡ＋Ｂ、Ｕ２＋Ａ及Ｕ１＋Ｃ对１４个血清型的检出率分别为１００％、８６％、７８％、６４％及６４％。结论引物自由能谱之差影响着ＰＣＲ扩增的敏感性。
Objective To compare the detecting rate and sensitivity of PCR by using different primers. Methods Five pairs of primers were designed according to the gene encoding urease for detecting Ureaplasma urealyticum(UU).The relationship between PCR sensitivity and primer sequences was analysed by software OLIGO. Results Different primers had different reactivity for serovar 1-14 of UU,and distinct sensitivity for detecting UU DNA.The difference of the sensitivity of different primers was as high as 40 folds. Conclusions Analysis of a variety of primer parameters indicated that the spectrum of free energy(ΔG) of primer pairs was decisive for PCR sensitivity.Detection of UU from clinical specimens demonstrated that only one primer pairs(1+B) of the four could detect all the serovars of UU in the specimens.
关键词
解脲脲原体
聚合酶链反应
引物
血清型
Ureaplasma urealyticum Polymerase chain reaction Primer Specimens
