中华皮肤科杂志 ›› 1994, Vol. 27 ›› Issue (3): 150-151,190.

• 论著 • 上一篇    下一篇

聚合酶链反应检测解脲支原体

张树文1, 王荷英2, 叶顺章2, 杨瑞馥3, 郭兆彪3, 李银太3   

  1. 1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042;
    2. 指导者;
    3. 军事医学科学院微生物流行病研究所
  • 收稿日期:1993-10-11 修回日期:1993-12-28 出版日期:1994-06-15 发布日期:1994-06-15

Detection of Ureaplasma Urealyticum Using the Polymerase Chain Reaction

ZHANG Shu-Wen1, WANG He-Ying2, YE Shun-Zhang2   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042
  • Received:1993-10-11 Revised:1993-12-28 Online:1994-06-15 Published:1994-06-15

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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.

关键词: 解脲支原体, 聚合酶链反应

Abstract: A polymerase chain reaction(PCR) assay was developed 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 demonstrated a sensitivity of 10-"g of DNA,which corresponds 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 sufficiently simple for routine application,may prove to be a valuable diagnostic tool.

Key words: Ureaplasma urealyticum, Polymerase chain reaction