DEVELOPMENT OF A PCR PROBE TEST FOR IDENTIFYING PSEUDOMONAS-AERUGINOSA AND PSEUDOMONAS (BURKHOLDERIA) CEPACIA

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 (Burkhold eria) cepacia in sputum from children with cystic fibrosis. Methods-Mo st of the 16s rRNA sequences from strains of Ps aeruginosa, Ps (Burkho lderia) cepacia, and Ps putida were determined. PCR primers and DNA pr obes were synthesised from suitable sequences and then evaluated on ba cterial cultures and sputum samples. Results-About 1000 bases of seque nce was obtained from strains of Ps aeruginosa, Ps (Burkholderia) cepa cia, and Ps putida. PCR of bacterial cultures was species specific, bu t PCR on sputum resulted in some non-specific amplification products. The subsequent hybridisation reaction was species specifc. Conclusion- A species specific system of PCR and DNA hybridisation based on 16s rR NA sequences is applicable in clinical practice, and may aid the early diagnosis of respiratory tract infection with small numbers of Ps aer uginosa and Ps (Burkholderia) cepacia in patients with cystic fibrosis .