肽核酸-荧光原位杂交快速鉴定化妆品中的铜绿假单胞菌

Rapid identification of Pseudomonas aeruginosa by peptide nucleic acid fluorescence in-situ hybridization

目的建立一种化妆品中铜绿假单胞菌(Pseudomonas aeruginosa)的快速检测方法。方法基于铜绿假单胞菌的16S r RNA序列设计铜绿假单胞菌特异性肽核酸(peptide nucleic acid,PNA)探针PA-16S-1,并对其探针进行灵敏度和特异性验证,建立并优化固相肽核酸荧光原位杂交(peptide nucleic acid-fluorescence in situ hybridization,PNA-FISH)检测方法。结果采用PNA-FISH法对300份化妆品样本进行检测,检测到2株铜绿假单胞菌和1株金黄色葡萄球菌阳性样品,与传统生化方法结果完全一致。结论 PNA-FISH方法是一种快速、敏感、特异的致病菌检测技术,对控制和提高化妆品的卫生质量具有重要意义。

Objective To establish a method for rapid detection of Pseudomonas aeruginosa in cosmetics.Methods The Pseudomonas aeruginosa species-specific peptide nucleic acid （PNA） probe PA-16S-1 was designed based on the conserved regions in 16S rRNA sequences,and its specificity and sensitivity were experimentally verified.The peptide nucleic acid fluorescence in-situ hybridization （PNA-FISH） method was developed for specific detection of Pseudomonas aeruginosa.Results Three hundred cosmetic samples were detected by PNA-FISH,2 strains of Pseudomonas aeruginosa and 1 strain of Staphylococcus aureus positive samples were identified,which were exactly the same as the results of traditional biochemical identification.Conclusion The established PNA-FISH identification method provides a fast,sensitive,and specific detection of pathogenic bacteria in cosmetics,which has an important significance to improve the sanitary quality of cosmetics.