Scholarlink

<p><i>Acetilactobacillus jinshanensis</i> (<i>A. jinshanensis</i>) is a key microorganism in solid-state vinegar fermentation, and it considerably contributes to the quality of vinegar. However, its growth in vinegar can lead to swollen deterioration, posing a serious threat to vinegar producers. In this study, a PMA-qPCR assay was developed to monitor <i>A. jinshanensis</i> during production. The specific primers targeting the <i>recA</i> gene were designed, and propidium monoazide (PMA) demonstrating the ability to differentiate between viable and dead bacteria was used. The optimal conditions for PMA action were determined as 2&#xa0;µg/mL PMA, 10&#xa0;min incubation in darkness, and 10&#xa0;min photoreaction. Under these conditions, minimal impact on viable cells and maximum signal reduction for dead cells were achieved. Compared with the qPCR method, the PMA-qPCR method could effectively inhibit 99% of dead cells, with recoveries ranging from 95 to 105%. The developed PMA-qPCR method demonstrated high sensitivity and specificity for detecting <i>A. jinshanensis</i>, with the ability to detect the microorganism within 2–3&#xa0;h. PMA-qPCR was employed to investigate the contamination sources of <i>A. jinshanensis</i> responsible for vinegar-swollen deterioration. The method demonstrated a strong correlation with traditional culture techniques. Results revealed that <i>A. jinshanensis</i> infiltrated the vinegar through the leaching process, remained insufficiently inactivated during the boiling process, and subsequently proliferated in aging tanks, ultimately leading to swollen deterioration. This study provides insights for vinegar production and risk management.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Development and Application of PMA-qPCR Based Methodologies on Viable Cell Quantification of Acetilactobacillus jinshanensis in Traditional Chinese Vinegar

  • 期刊论文

  • 发表日期：

    2025年8月22日

查看全文
  • Kui Zhao,
  • Kaidi Hu,
  • Xinyue Gu,
  • Jianlong Li,
  • Qin Li,
  • Ning Zhao,
  • Xinjie Hu,
  • Aiping Liu,
  • Shuliang Liu

摘要

Acetilactobacillus jinshanensis (A. jinshanensis) is a key microorganism in solid-state vinegar fermentation, and it considerably contributes to the quality of vinegar. However, its growth in vinegar can lead to swollen deterioration, posing a serious threat to vinegar producers. In this study, a PMA-qPCR assay was developed to monitor A. jinshanensis during production. The specific primers targeting the recA gene were designed, and propidium monoazide (PMA) demonstrating the ability to differentiate between viable and dead bacteria was used. The optimal conditions for PMA action were determined as 2 µg/mL PMA, 10 min incubation in darkness, and 10 min photoreaction. Under these conditions, minimal impact on viable cells and maximum signal reduction for dead cells were achieved. Compared with the qPCR method, the PMA-qPCR method could effectively inhibit 99% of dead cells, with recoveries ranging from 95 to 105%. The developed PMA-qPCR method demonstrated high sensitivity and specificity for detecting A. jinshanensis, with the ability to detect the microorganism within 2–3 h. PMA-qPCR was employed to investigate the contamination sources of A. jinshanensis responsible for vinegar-swollen deterioration. The method demonstrated a strong correlation with traditional culture techniques. Results revealed that A. jinshanensis infiltrated the vinegar through the leaching process, remained insufficiently inactivated during the boiling process, and subsequently proliferated in aging tanks, ultimately leading to swollen deterioration. This study provides insights for vinegar production and risk management.