中华皮肤科杂志 ›› 1998, Vol. 31 ›› Issue (3): 160-162.

• 论著 • 上一篇    下一篇

脉冲场凝胶电泳及随机扩增多态性DNA对淋球菌的基因分型

苏晓红, 叶顺章   

  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:1997-07-06 修回日期:1997-10-29 出版日期:1998-06-15 发布日期:1998-06-15

Molecular Typing of Neisseria Gonorrhoeae by Pulsed field Gel Electrophoresis and Random Amplified Polymorphic DNA Analysis

Su Xiaohong, Ye Shunzhang   

  1. Institute of Dermatology, Chinese Academy of Medical Scienecs, Peking Union Medical College, Nanjing 210042
  • Received:1997-07-06 Revised:1997-10-29 Online:1998-06-15 Published:1998-06-15

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摘要： 目的 评价脉冲场凝胶电泳限制性内切酶分析(PFGEREA)及随机扩增多态性DNA(RAPD)分析对淋球菌分离株进行基因分型的能力.方法 36株淋球菌分离株基因组DNA以SpeⅠ酶切及脉冲场凝胶电泳.并以随机引物OPA-03对36株菌进行RAPD分析.结果 PFGEREA在36株菌中产生12～19个DNA片段(5～600kb),区分出20种PFGE型.以OPA-03扩增淋球菌DNA,获得15个不同的DNA片段(280～1900bp).每株菌有3～9个片段.36株菌呈现18种RAPD型.研究表明,PFGEREA与RAPD可区分属相同营养型及抗生素谱的菌株,二种方法分型结果总的趋向一致.不同地区的分离株甚至同一地区的某些分离株存在相当大的DNA多态性.具相同或十分相似基因型的菌株为某一特定地区所特有.认为PFGEREA及RAPD是鉴别淋球菌临床分离株有用而敏感的分子技术,有助于了解菌株来源、菌株间的克隆相关性及抗生素耐药性的传播.与PFGEREA相比,RAPD分析相对快速、简便和经济.

关键词: 淋球菌, 基因分型, PFGE, RAPD

Abstract: Objective To evaluate the ability of restriction endonuclease analysis by pulsed field gel electrophoresis (PFGE REA) and random amplified polymorphic DNA (RAPD) analysis for molecular typing of Neisseria gonorrhoeae isolates.Methods Genomic DNA from 36 Neisseria gonorrhoeae isolates were digested with SpeI and analysed by contour clamped homogeneous electric fields electrophoresis.The 36 isolates were also characterised by RAPD analysis with arbitrary primer OPA 03.Results 12～19 DNA fragments from 5 to 600kb were obtained among the 36 isolates after PFGE REA.The 36 isolates were discriminated into 20 PFGE patterns.Amplification of Neisseria gonorrhoeae DNA with primer OPA 03 produced 15 different DNA fragments (280～1900bp),and 3～9 fragments per strain could be seen.The 36 isolates showed 18 different RAPD patterns.Strains sharing common auxotypes and antibiotic spectrum could be differentiated by PFGE REA and RAPD analysis.General agreement was found between these two techniques.Isolates from different geographic areas and even some isolates in the same area showed considerable amount of DNA polymorphism.In addition,some isolates shared common or very similar patterns were unique to a given geographic area.Conclusion It is concluded that both PFGE REA and RAPD analysis are useful,sensitive molecular techniques for differentiation of clinical isolates of Neisseria gonorrhoeae.They should be helpful in the investigation of strain origin,clonal relation among strains and spread of antibiotic resistance.Compared with PFGE REA,RAPD analysis is faster,relatively simple and more economical.

Key words: Neisseria Gonorrhoeae, Genotyping, PFGE, RAPD