RRC ID 71067
著者 Niikura M, Atobe S, Takahashi A, Kado Y, Sugimoto T, Tsuji H, Shimizu K, Ogura H, Asahara T.
タイトル Development of a rapid and sensitive analytical system for Pseudomonas aeruginosa based on reverse transcription quantitative PCR targeting of rRNA molecules.
ジャーナル Emerg Microbes Infect
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.
巻・号 10(1)
ページ 677-686
公開日 2021-12-1
DOI 10.1080/22221751.2021.1906164
PMID 33734032
PMC PMC8023615
MeSH Adult Aged Aged, 80 and over Anti-Bacterial Agents / therapeutic use Blood / microbiology Caco-2 Cells DNA, Bacterial Diagnostic Tests, Routine / methods* Drug Resistance, Bacterial Early Diagnosis Feces / microbiology Female Humans Infection Control / methods Intensive Care Units Male Microbiota Middle Aged Molecular Diagnostic Techniques / methods* Polymerase Chain Reaction Pseudomonas Infections / diagnosis* Pseudomonas Infections / microbiology Pseudomonas aeruginosa / classification* Pseudomonas aeruginosa / drug effects Pseudomonas aeruginosa / genetics* RNA, Ribosomal, 23S Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Young Adult
リソース情報
一般微生物 JCM2776 JCM5961 JCM2952 JCM3843 JCM5968 JCM1471 JCM1112 JCM1152