Department of Clinical Oral Oncology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Department of Oral Health, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
J Microbiol Methods. 2023 Nov;214:106844. doi: 10.1016/j.mimet.2023.106844. Epub 2023 Oct 17.
Aspiration pneumonia can occur in perioperative and older patients, and various oral care methods have been used to prevent it. To validate the effective oral care methods, measuring bacterial counts before and after oral care is necessary. However, isolating and quantifying viable bacteria from those that are inactivated by agents used in oral care is not possible. In this study, we developed a novel method, Delayed real-time PCR (DR-PCR), that can quantify only viable bacteria from mixed samples of viable and dead bacteria. This method takes advantage of the fact that dead bacteria do not grow but viable bacteria do. When the samples were incubated in a liquid medium for 4 hours, the higher the percentage of viable bacteria, the higher the rate of increase in the number of bacteria. This method showed that povidone‑iodine mouthwashing reduced the number of viable bacteria to approximately 1/4 of that before mouthwashing. Although DR-PCR is slightly more time consuming than real-time PCR, it is effective for studying changes in bacterial counts before and after oral care.
吸入性肺炎可发生于围手术期和老年患者，已采用多种口腔护理方法来预防其发生。为了验证有效的口腔护理方法，有必要在口腔护理前后测量细菌计数。然而，用口腔护理剂灭活的细菌与活细菌混合时，无法对其进行分离和定量。在本研究中，我们开发了一种新方法，即延迟实时 PCR（DR-PCR），该方法仅能对活细菌与死细菌的混合样本中的活细菌进行定量。该方法利用了这样一个事实，即死细菌不生长而活细菌会生长。当将样本在液体培养基中孵育 4 小时时，活细菌的百分比越高，细菌数量的增加率越高。该方法表明，聚维酮碘漱口可将活细菌数量减少到漱口前的约 1/4。虽然 DR-PCR 比实时 PCR 稍微耗时，但它对于研究口腔护理前后细菌计数的变化非常有效。
