Kur J, Burkiewicz A, Zablocka A, Gospodarek E
Department of Microbiology, Technical University of Gdansk, Poland.
Acta Microbiol Pol. 1995;44(2):111-7.
In the present study, 40 clinical strains of Pseudomonas aeruginosa were investigated by using the polymerase chain reaction (PCR). We used primers to amplify spacer regions between the 16S and 23S genes in the prokaryotic rRNA genetic loci. When the spacer amplification products were resolved by electrophoresis, the resulting patterns were characteristic for all tested strains. Only one specific PCR fragment of 580 bp was formed. This product was digested with HaeIII, HinfI and AluI restriction endonucleases. Restriction fragments produced by all three restriction endonucleases were characteristic and have the same sizes for all strains tested. The amplification product contains a conserved, internal single HaeIII restriction site. On the basis of our results, PCR amplifications of the 16S-23S spacer region for Pseudomonas aeruginosa and subsequent RFPL analysis show significant promise as a tool for the simple identification of this bacteria.
在本研究中，我们使用聚合酶链反应（PCR）对40株铜绿假单胞菌临床菌株进行了研究。我们使用引物扩增原核rRNA基因位点中16S和23S基因之间的间隔区。当间隔区扩增产物通过电泳分离时，所得图谱对所有测试菌株具有特征性。仅形成了一个580 bp的特异性PCR片段。该产物用HaeIII、HinfI和AluI限制性内切酶进行消化。所有三种限制性内切酶产生的限制性片段具有特征性，并且对所有测试菌株大小相同。扩增产物包含一个保守的内部单一HaeIII限制性位点。基于我们的结果，铜绿假单胞菌16S - 23S间隔区的PCR扩增及随后的限制性片段长度多态性（RFLP）分析显示，作为一种简单鉴定该细菌的工具具有很大潜力。
