用于鉴定铜绿假单胞菌和洋葱伯克霍尔德菌（假单胞菌属）的聚合酶链式反应（PCR）探针检测法的开发。
Development of a PCR probe test for identifying Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia.
作者信息
O'Callaghan E M, Tanner M S, Boulnois G J
机构信息
Department of Child Health, University of Leicester.
出版信息
J Clin Pathol. 1994 Mar;47(3):222-6. doi: 10.1136/jcp.47.3.222.
AIMS
To develop a system of species specific polymerase chain reaction (PCR) and DNA hybridisation based on 16s ribosomal RNA sequences for the identification of Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia in sputum from children with cystic fibrosis.
METHODS
Most of the 16s rRNA sequences from strains of Ps aeruginosa, Ps (Burkholderia) cepacia, and Ps putida were determined. PCR primers and DNA probes were synthesised from suitable sequences and then evaluated on bacterial cultures and sputum samples.
RESULTS
About 1000 bases of sequence was obtained from strains of Ps aeruginosa, Ps (Burkholderia) cepacia, and Ps putida. PCR of bacterial cultures was species specific, but PCR on sputum resulted in some non-specific amplification products. The subsequent hybridisation reaction was species specific.
CONCLUSION
A species specific system of PCR and DNA hybridisation based on 16s rRNA sequences is applicable in clinical practice, and may aid the early diagnosis of respiratory tract infection with small numbers of Ps aeruginosa and Ps (Burkholderia) cepacia in patients with cystic fibrosis.
目的
开发一种基于16s核糖体RNA序列的物种特异性聚合酶链反应（PCR）和DNA杂交系统，用于鉴定囊性纤维化患儿痰液中的铜绿假单胞菌和洋葱伯克霍尔德菌（假单胞菌属）。
方法
测定了铜绿假单胞菌、洋葱伯克霍尔德菌（假单胞菌属）和恶臭假单胞菌菌株的大部分16s rRNA序列。从合适的序列合成PCR引物和DNA探针，然后在细菌培养物和痰液样本上进行评估。
结果
从铜绿假单胞菌、洋葱伯克霍尔德菌（假单胞菌属）和恶臭假单胞菌菌株中获得了约1000个碱基的序列。细菌培养物的PCR具有物种特异性，但痰液的PCR产生了一些非特异性扩增产物。随后的杂交反应具有物种特异性。
结论
基于16s rRNA序列的物种特异性PCR和DNA杂交系统适用于临床实践，可能有助于早期诊断囊性纤维化患者中少量铜绿假单胞菌和洋葱伯克霍尔德菌（假单胞菌属）引起的呼吸道感染。
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