Establishment of a rapid detection method for Pseudomonas aeruginosa based on real-time fluorescent recombinase polymerase amplification technology
10.3969/j.issn.1673-4130.2024.19.005
- VernacularTitle:基于实时荧光重组酶聚合酶扩增技术的铜绿假单胞菌快速检测方法的建立
- Author:
Yun XING
1
;
Yan ZHANG
;
Daohong ZHOU
;
Qiu ZHONG
;
Yuansu JIANG
;
Qing HUANG
;
Baihui ZHENG
Author Information
1. 中国人民解放军陆军特色医学中心检验科,重庆 400042
- Keywords:
Pseudomonas aeruginosa;
exoS gene;
real-time fluorescent recombinase polymerase ampli-fication
- From:
International Journal of Laboratory Medicine
2024;45(19):2329-2333
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a real-time fluorescent recombinase polymerase amplification(RPA)technology for the detection of the virulence gene exoS of Pseudomonas aeruginosa,and evaluate the specifici-ty,sensitivity and practicability of the method.Methods According to the specific conserved region of the vir-ulence gene exoS of Pseudomonas aeruginosa,the specific primers and probes of RPA were designed,and the extracted target DNA was detected to determine the specificity and sensitivity of RPA.Real-time fluorescence quantitative PCR(qPCR)was established to detect the target DNA,and the detection limits of different detec-tion methods for Pseudomonas aeruginosa were compared.The feasibility of RPA in detecting Pseudomonas aeruginosa was further confirmed by the performance verification test of clinical samples.Results The estab-lished RPA detection method had good specificity.Only Pseudomonas aeruginosa had specific amplification curve,but no specific amplification curve for other bacteria.The sensitivity of RPA was 5×102 cfu/mL,which was consistent with the detection limit of qPCR and the results were reliable.The detection time of RPA method was only 30 min,which was significantly lower than that of the traditional method.Conclusion The RPA method for the detection of Pseudomonas aeruginosa established in this study has high specificity and sensitivity,and significantly shortens the detection time compared with the traditional detection method.It can be used for the rapid detection of Pseudomonas aeruginosa in clinical specimens.
