聚合酶链反应检测杜克雷嗜血杆菌DNA
Polymerase chain reaction detection of Haemophilus ducreyi DNA.
作者信息
Roesel D J, Gwanzura L, Mason P R, Joffe M, Katzenstein D A
机构信息
Center for AIDS Research, Stanford University, California 94305-5107, USA.
出版信息
Sex Transm Infect. 1998 Feb;74(1):63-5. doi: 10.1136/sti.74.1.63.
OBJECTIVES
To develop a polymerase chain reaction (PCR) method to detect Haemophilus ducreyi DNA in cultured isolates and clinical material.
METHODS
Primers specific to the H ducreyi 16s rRNA gene were synthesised. PCR conditions were optimised and products were verified by restriction endonuclease digestion and agarose gel electrophoresis.
RESULTS
The method was able to detect all 28 H ducreyi strains tested; specificity was demonstrated using lysates of 12 related organisms. Applied to clinical samples from genital ulcer swabs obtained in Harare, Zimbabwe, H ducreyi DNA was detected in repeated assays in 35 clinical samples.
CONCLUSION
PCR amplification using primers from the 16s rRNA gene may be a useful alternative to culture for the detection of H ducreyi and the diagnosis of chancroid.
目的
开发一种聚合酶链反应（PCR）方法，用于检测培养分离株和临床材料中的杜克雷嗜血杆菌DNA。
方法
合成针对杜克雷嗜血杆菌16s rRNA基因的引物。优化PCR条件，并通过限制性内切酶消化和琼脂糖凝胶电泳验证产物。
结果
该方法能够检测所有28株受试杜克雷嗜血杆菌菌株；使用12种相关生物体的裂解物证明了其特异性。应用于从津巴布韦哈拉雷获得的生殖器溃疡拭子的临床样本，在35份临床样本的重复检测中检测到了杜克雷嗜血杆菌DNA。
结论
使用来自16s rRNA基因的引物进行PCR扩增，可能是检测杜克雷嗜血杆菌和诊断软下疳的一种有用的替代培养方法。
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