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首页> 外文期刊>Food microbiology >Quantitative analysis of viable, stressed and dead cells of Campylobacter jejuni strain 81-176
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Quantitative analysis of viable, stressed and dead cells of Campylobacter jejuni strain 81-176

机译：空肠弯曲杆菌81-176株存活，应激和死细胞的定量分析

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摘要

Campylobacter jejuni is an important foodborne gastrointestinal pathogen and highly sensitive to environmental stresses. Research has shown that changes in culturability, cell morphology, and viability occur when C. jejuni cells are subjected to stresses. In this study, real-time PCR, ethidium monoazide (EMA) in combination with real-time PCR (EMA-PCR), BacLight bacterial viability staining, and agar plate counting methods were used to quantitatively analyze viable, stressed, and dead C. jejuni strain 81-176. The real-time PCR assay provides highly sensitive and specific quantification of total genome copies of C. jejuni culture in different growth phases. Our results also reveal that real-time PCR can be used for direct quantification of Campylobacter genome release into Phosphate Buffered Saline (PBS) as an indicator of cell lysis. Using EMA-PCR, we obtained a dynamic range of greater than 3 logs for differentiating viable vs. dead cells. The viability and morphological characteristics of the stressed cells after one-week incubation at 25 ℃, in air, and under nutrient-poor conditions were investigated. Our results indicated that, over 99% of the stressed cells were converted from the spiral to the coccoid form and became non-culturable. However, more than 96% of the coccoid cells retained their membrane integrity as suggested by both the BacLight staining and EMA-PCR analyses. Thus, to detect C. jejuni under stress conditions, conventional culturing method in conjunction with EMA-PCR or BacLight staining might be a more appropriate approach.
机译：空肠弯曲菌是一种重要的食源性胃肠道病原体，对环境压力高度敏感。研究表明，当空肠弯曲杆菌细胞受到压力时，可培养性，细胞形态和生存能力就会发生变化。在这项研究中，使用实时PCR，单叠氮化乙锭（EMA）结合实时PCR（EMA-PCR），BacLight细菌生存力染色和琼脂平板计数方法来定量分析存活，受压和死亡的C。空肠菌株81-176。实时PCR分析可对不同生长阶段空肠弯曲菌培养的总基因组拷贝进行高度灵敏和特异性的定量。我们的结果还表明，实时PCR可以用于弯曲杆菌基因组释放到磷酸盐缓冲盐水（PBS）中作为细胞裂解的指标的直接定量。使用EMA-PCR，我们获得了大于3个对数的动态范围，用于区分存活细胞与死细胞。研究了25℃，在空气中和营养不良条件下培养1周后应激细胞的活力和形态特征。我们的结果表明，超过99％的应激细胞从螺旋形转变为类球体形式，变得不可培养。然而，如BacLight染色和EMA-PCR分析所表明的，超过96％的类球菌细胞保留了其膜的完整性。因此，为了在压力条件下检测空肠弯曲杆菌，常规培养方法结合EMA-PCR或BacLight染色可能是更合适的方法。

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  • 来源
    《Food microbiology》 |2010年第4期|p.439-446|共8页
  • 作者

    Yiping He; Chin-Yi Chen;

  • 作者单位

    Microbial Biophysics and Residue Chemistry Research Unit, U. S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA;

    Microbial Food Safety Research Unit, Eastern Regional Research Center, Agricultural Research Service, U. S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    campylobacter jejuni; ethidium monoazide; real-time PCR; viability; stressed cells;

    机译：空肠弯曲菌单叠氮化乙锭;实时PCR;可行性;压力细胞;

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