Yakult Central Institute, Yakult Honsha Co., Ltd., Kunitachi, Tokyo, Japan.
Department of Traumatology and Acute Critical Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Emerg Microbes Infect. 2021 Dec;10(1):677-686. doi: 10.1080/22221751.2021.1906164.
For (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%, < 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.
对于（PA），感染控制和适当的抗菌治疗已成为重要问题。诊断对于管理 PA 感染至关重要，但常规方法的准确性和速度都不高。我们开发了一种基于 23S rRNA 靶向逆转录定量 PCR（RT-qPCR）的新型 PA 定量系统。我们证实 RT-qPCR 可以快速（6 小时内）直接从临床样本中定量 PA，具有高灵敏度（血液为 1 个细胞/mL；粪便为 100 个细胞/g）且无交叉反应。此外，在抗生素治疗下，与培养和 qPCR 等其他方法相比，该系统检测到的 PA 活菌数与感染的 Caco-2 细胞的炎症反应相关性更好。接下来，我们在从 65 名脓毒症 ICU 患者和 44 名健康志愿者收集的粪便样本上使用该系统来识别 ICU 感染状态。我们证实 ICU 患者的 PA 检测率明显高于健康志愿者（49.2%比 13.6%，<0.05）。此外，我们通过该系统监测了 4 名 ICU 患者的耐药 PA。PA 计数的趋势准确反映了各种治疗背景，如抗生素使用和机械通气使用。我们的结果表明，该 RT-qPCR 系统有助于早期诊断和评估适当的抗菌治疗，可能是对抗 PA 感染的有用工具。
