不同DNA提取方法对PCR检测解脲支原体的影响

Comparison of three different DNA extraction methods affecting the results of PCR detection for Ureaplasma urealyticum

目的 : 比较PK、NK和PCI三种不同DNA提取方法对解脲支原体PCR的影响。方法 : 从临床得到的 330份怀孕妇女的尿标本 ,接种到液体培养基和A8琼脂平板 ,液体培养阳性的标本 1 10稀释后 ,用PK、NK和PCI三种不同DNA提取方法进行提取 ,分别作PCR及抑制实验 ,比较三种提取方法的敏感性和特异性。结果 : 液体培养和A8琼脂平板培养的阳性率分别为 6 8.1%和 39%,两者符合率为 76 %,PK、NK和PCI的敏感性为 5 2 .5 %,5 5 %,87.5 %;特异性为 6 5 .8%,76 .3%,6 0 .5 %;抑制率为 33.3%,33.0 %,3.3%。结论 : 在PK、NK和PCI三种不同DNA提取方法中 ,以PCI敏感性最高 ,抑制率最低。

Objective:In order to compare three different DNA extraction methods, which affect the results of PCR detection for Ureaplasma urealyticum.Method: Urine specimens from 330 pregnant women were inoculated into the ureaplasma broth and A8 agar plates. After the positively cultured ureaplasma-broth specimens were diluted at 1:10, the template DNAs were extracted by PK, NK and PCI methods. And then, amplification and inhibition tests by PCR were performed, and the sensitivity and specificity of the tests were compared. Results:The positivity rates of ureaplasma-broth and A8 agar plate were 68.1% and 39% respectively. The agreement rate was 76%. The sensitivity of PK, NK and PCI was 52.5%, 55.0% and 87.5%, respectively, while the specificity of the three methods was 65.8%, 76.3% and 60.5%, respectively. The inhibition rates in the three methods were 33.3%, 3 3.0% and 3.3% respectively. Conclusion: PCI method was the most sensitive with the lowest inhibition rate among the three different DNA extraction methods.