{"title":"[PMA-ddPCR法检测可存活但不可培养的高乙醇产肺炎克雷伯菌]。","authors":"S Zhao, C P Dou, J Zhang, J Yuan","doi":"10.3760/cma.j.cn112150-20240401-00265","DOIUrl":null,"url":null,"abstract":"<p><p><b>Objective:</b> To establish an absolute quantitative method for high ethanol-producing <i>klebsiella pneumoniae</i> in a viable non-culturable (VBNC) state. <b>Methods:</b> High ethanol-producing <i>Klebsiella pneumonia</i> was induced to enter the VBNC state and then the ethanol production was evaluated. A PMA-ddPCR method was established to count the copies of live cell genes in the VBNC state of high ethanol-producing <i>Klebsiella pneumoniae</i> using single-copy genes. Further, the sensitivity and adaptability of ddPCR for detecting low-concentration samples were evaluated in VBNC fecal simulation. <b>Results:</b> The lower detection limit of ddPCR for quantitative analysis of high ethanol-producing <i>Klebsiella pneumoniae</i> gradient diluent was 10 times that of qPCR. At low temperature and low nutritional state, high ethanol-producing <i>Klebsiella pneumoniae</i> entered the VBNC state on the 45<sup>th</sup> day. The quantitative results of PMA-ddPCR on VBNC state cells were (5.46±0.05) log<sub>10</sub> DNA copies/ml. The ethanol production in the VBNC state was<2.2 mmol/L and the ability to produce ethanol was restored after recovery. The minimum detection limit for ddPCR in fecal simulated samples with VBNC state was 3.2 log<sub>10</sub> DNA copies/ml. <b>Conclusion:</b> The ddPCR detection method for high ethanol-producing <i>Klebsiella pneumoniae</i> with VBNC state has good sensitivity and adaptability, and can be used for the detection of VBNC state cells in clinical samples.</p>","PeriodicalId":24033,"journal":{"name":"中华预防医学杂志","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[PMA-ddPCR method for detecting high ethanol-producing <i>klebsiella pneumoniae</i> in viable but non-culturable state].\",\"authors\":\"S Zhao, C P Dou, J Zhang, J Yuan\",\"doi\":\"10.3760/cma.j.cn112150-20240401-00265\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Objective:</b> To establish an absolute quantitative method for high ethanol-producing <i>klebsiella pneumoniae</i> in a viable non-culturable (VBNC) state. <b>Methods:</b> High ethanol-producing <i>Klebsiella pneumonia</i> was induced to enter the VBNC state and then the ethanol production was evaluated. A PMA-ddPCR method was established to count the copies of live cell genes in the VBNC state of high ethanol-producing <i>Klebsiella pneumoniae</i> using single-copy genes. Further, the sensitivity and adaptability of ddPCR for detecting low-concentration samples were evaluated in VBNC fecal simulation. <b>Results:</b> The lower detection limit of ddPCR for quantitative analysis of high ethanol-producing <i>Klebsiella pneumoniae</i> gradient diluent was 10 times that of qPCR. At low temperature and low nutritional state, high ethanol-producing <i>Klebsiella pneumoniae</i> entered the VBNC state on the 45<sup>th</sup> day. The quantitative results of PMA-ddPCR on VBNC state cells were (5.46±0.05) log<sub>10</sub> DNA copies/ml. The ethanol production in the VBNC state was<2.2 mmol/L and the ability to produce ethanol was restored after recovery. The minimum detection limit for ddPCR in fecal simulated samples with VBNC state was 3.2 log<sub>10</sub> DNA copies/ml. <b>Conclusion:</b> The ddPCR detection method for high ethanol-producing <i>Klebsiella pneumoniae</i> with VBNC state has good sensitivity and adaptability, and can be used for the detection of VBNC state cells in clinical samples.</p>\",\"PeriodicalId\":24033,\"journal\":{\"name\":\"中华预防医学杂志\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-07-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华预防医学杂志\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3760/cma.j.cn112150-20240401-00265\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华预防医学杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3760/cma.j.cn112150-20240401-00265","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
[PMA-ddPCR method for detecting high ethanol-producing klebsiella pneumoniae in viable but non-culturable state].
Objective: To establish an absolute quantitative method for high ethanol-producing klebsiella pneumoniae in a viable non-culturable (VBNC) state. Methods: High ethanol-producing Klebsiella pneumonia was induced to enter the VBNC state and then the ethanol production was evaluated. A PMA-ddPCR method was established to count the copies of live cell genes in the VBNC state of high ethanol-producing Klebsiella pneumoniae using single-copy genes. Further, the sensitivity and adaptability of ddPCR for detecting low-concentration samples were evaluated in VBNC fecal simulation. Results: The lower detection limit of ddPCR for quantitative analysis of high ethanol-producing Klebsiella pneumoniae gradient diluent was 10 times that of qPCR. At low temperature and low nutritional state, high ethanol-producing Klebsiella pneumoniae entered the VBNC state on the 45th day. The quantitative results of PMA-ddPCR on VBNC state cells were (5.46±0.05) log10 DNA copies/ml. The ethanol production in the VBNC state was<2.2 mmol/L and the ability to produce ethanol was restored after recovery. The minimum detection limit for ddPCR in fecal simulated samples with VBNC state was 3.2 log10 DNA copies/ml. Conclusion: The ddPCR detection method for high ethanol-producing Klebsiella pneumoniae with VBNC state has good sensitivity and adaptability, and can be used for the detection of VBNC state cells in clinical samples.
期刊介绍：
Chinese Journal of Preventive Medicine (CJPM), the successor to Chinese Health Journal , was initiated on October 1, 1953. In 1960, it was amalgamated with the Chinese Medical Journal and the Journal of Medical History and Health Care , and thereafter, was renamed as People’s Care . On November 25, 1978, the publication was denominated as Chinese Journal of Preventive Medicine . The contents of CJPM deal with a wide range of disciplines and technologies including epidemiology, environmental health, nutrition and food hygiene, occupational health, hygiene for children and adolescents, radiological health, toxicology, biostatistics, social medicine, pathogenic and epidemiological research in malignant tumor, surveillance and immunization.
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