中华皮肤科杂志 ›› 2003, Vol. 36 ›› Issue (12): 679-681.

• 论著 • 上一篇    下一篇

鸟分支杆菌聚合酶链反应检测的研究

李晓杰, 吴勤学, 刘训荃   

  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
  • 收稿日期:2002-12-25 出版日期:2003-12-15 发布日期:2003-12-15
  • 通讯作者: 吴勤学

Study on Detection of Mycobacterium avium by Polymerase Chain Reaction

LI Xiao-jie, WU Qin-xue, LIU Xun-quan   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China
  • Received:2002-12-25 Online:2003-12-15 Published:2003-12-15

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摘要： 目的 建立敏感性高、特异性强的快速检测鸟分支杆菌的方法.方法 用特异性引物对连续稀释的鸟分支杆菌菌悬液DNA进行PCR扩增,验证其敏感性;并用该引物对除鸟分支杆菌外的其他17株分支杆菌DNA进行扩增,验证其特异性;将正常皮肤组织与鸟分支杆菌的菌悬液混合以模拟临床皮肤感染标本,初步探讨适宜的临床标本前处理的方法,以提高PCR方法在检测临床皮肤组织感染中的敏感性.结果 PCR方法可扩增427bp的鸟分支杆菌DNA片段,敏感性达1×102个菌细胞/mL,对其他分支杆菌进行扩增,结果均为阴性.对模拟临床皮肤感染标本进行PCR检测,敏感性降低为1×104个菌细胞/mL;将组织匀浆连续倍比稀释后,再加入细菌悬液,结果当组织匀浆稀释度≥1:4时,PCR的敏感性提高到1×102个菌细胞/mL.结论 PCR方法能敏感、特异地快速检测鸟分支杆菌.

关键词: 分枝杆菌,鸟, 聚合酶链反应

Abstract: Objective To develop a rapid method with high sensitivity and specificity to detect Mycobacterium avium.Methods A polymerase chain reaction(PCR)was developed.Its sensitivity and specificity were verified by sequential dilution of DNA of M.avium and other 17 Mycobacterium spp.,respectively.Simulation of clinical infection was established by mixing normal skin tissue with M.avium.The treatment of the tissue specimens was optimized in order to improve the sensitivity of the PCR assay.Results A fragment of 427bp was amplified by the PCR assay with the strains of M.avium at the sensitivity of 1×102 cells/mL.The other 17 Mycobacterium spp.were all negative.The sensitivity of the PCR assay decreased to 1×104cells/mL when M.avium was mixed with the homogenized skin tissue.The sensitivity recovered to 1×102 cell/mL when the skin tissue was diluted to≥1:4.Conclusion It is suggested that PCR be a rapid and reliable method for detection of M.avium.

Key words: Mycobacterium avium, Polymerase chain reaction