摘要
我们根据已知的解脲支原体尿素酶基因序列,设计了一对聚合酶链反应引物。该组引物对解脲支原体14个血清型均能扩增出283bp片段,但对实验所用的其它支原体或细菌不能扩增出任何片段。该283bp片段能被限制性内切酶Hinc Ⅱ降解为151,132bp两条片段。使用35个PCR循环,可检测出10-14g的模板DNA,约相当于20个解脲支原体。通过梯度稀释试验证明40个PCR循环时,PCR的敏感性与培养法的敏感性相当。用PCR检测解脲支原体操作简便,可作为一种有价值的诊断方法。
A polymerase chain reaction(PCR) assay was de- veloped for detection of Urea plasma urealyticum DNA. From the published sequence of the ureaplasma urease genes, two primers was selected' The primer sets amplified 283hp fragment from each of 14 serovars of ureasplasma,but did not amplify any fragment from other mycoplasmas or other oraganisms in this study.Those 283hp fragments could be digested to 151, 132hp fragments by Hinc Ⅱ. After 35 cycles of amplification with the primer sets,PCR assay demon- strated a sensitivity of 10--'g of DNA, which corre- sponds to the detection of 20 copies of Ureaplasma urealyticum. Serial dilution experiments revealed that the PCR procedure with 40 cycles of amplification was as sensitive as culture. The PCR assay, while suffi- ciently simple for routine application,may prove to be a valuable diagnostic tool.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
1994年第3期150-151,T003,共3页
Chinese Journal of Dermatology
关键词
解脲支原体
聚合酶链反应
Ureaplasma urealyticum Polymerase chain reaction