Development of a rapid and sensitive analytical system for Pseudomonas aeruginosa based on reverse transcription quantitative PCR targeting of rRNA molecules. Issue 1 (1st January 2021)

• Record Type: Journal Article
• Authors: Niikura, Mai; Atobe, Satomi; Takahashi, Akira; Kado, Yukiko; Sugimoto, Takuya; Tsuji, Hirokazu; Shimizu, Kentaro; Ogura, Hiroshi; Asahara, Takashi
• Is Part Of: Emerging microbes & infections. Volume 10:Issue 1(2021)
• Page Start: 677
• Page End: 686
• Publication Date: 2021-01-01
• Subjects: Pseudomonas aeruginosa -- infection -- RT-qPCR -- blood -- feces -- ICU patients -- antimicrobial resistance -- microbiota; Medical microbiology -- Periodicals; Communicable diseases -- Periodicals; Infection -- Periodicals; 616.9041
• DOI: 10.1080/22221751.2021.1906164
• Languages: English
• ISSNs: 2222-1751

Abstract

ABSTRACT: For Pseudomonas aeruginosa (PA), infection control and appropriate antimicrobial treatment have become important issues. Diagnosis is critical in managing PA infection, but conventional methods are not highly accurate or rapid. We developed a new PA quantification system based on 23S rRNA-targeted reverse transcription quantitative PCR (RT-qPCR). We confirmed that RT-qPCR can quantify PA directly from clinical samples quickly (within 6 h) and with high sensitivity (blood, 1 cell/mL; stool, 100 cells/g) and without cross-reaction. Also, under antibiotic treatment, PA viable counts detected by this system correlated well with the inflammatory response of infected Caco-2 cells compared to other methods such as culturing and qPCR. Next, we utilized this system on fecal samples collected from 65 septic ICU patients and 44 healthy volunteers to identify ICU infection status. We confirmed that the PA detection ratio in ICU patients was significantly higher than that in healthy volunteers (49.2% vs. 13.6%, P < 0.05). Additionally, we monitored drug-resistant PA in 4 ICU patients by this system. The trends in PA counts accurately reflected various treatment backgrounds such as antibiotic use and mechanical ventilator use. Our results suggest that this RT-qPCR system is beneficial for the early diagnosis and evaluation of appropriate antibacterial treatment and may be a useful tool in combating PA infection.