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通过 PCR 和荧光原位杂交技术特异性检测意大利南部河口水中的弯曲杆菌属。

Specific detection of Arcobacter spp. in estuarine waters of Southern Italy by PCR and fluorescent in situ hybridization.

机构信息

Dipartimento di Patologia e Microbiologia Sperimentale, Policlinico Universitario, Università di Messina, Messina, Italy.

出版信息

Lett Appl Microbiol. 2010 Jan;50(1):65-70. doi: 10.1111/j.1472-765X.2009.02767.x.

Abstract

AIM

To evaluate the reliability of culture-independent methods in comparison with culture-dependent ones for the detection of Arcobacter spp. in estuarine waters of Southern Italy.

METHODS AND RESULTS

PCR and fluorescent in situ hybridization (FISH) procedures were used to detect arcobacters directly in water samples and after enrichment cultures. The samples totally were positive by molecular methods (PCR and FISH) but only 75% were culture positive, confirming the limitation of these latter to detect Arcobacter spp. in natural samples. Culturable arcobacters were retrieved in all times except in July, and isolated species were ascribed only to Arcobacter cryaerophilus.

CONCLUSIONS

Culturable and nonculturable forms of Arcobacter in the estuarine environment were present. PCR assays were more sensitive than traditional culture in detecting Arcobacter butzleri and A. cryaerophilus. FISH comparatively to PCR technique may provide information about cell morphology and viability of single cells.

SIGNIFICANCE AND IMPACT OF THE STUDY

Our investigation indicates the existence of an environmental reservoir of potential pathogenic arcobacters in an estuarine Italian area, which may survive under a viable but not culturable state.

摘要

目的

评估与培养依赖方法相比,非培养方法在检测意大利南部河口水中弯曲菌属的可靠性。

方法与结果

PCR 和荧光原位杂交(FISH)程序用于直接在水样和富集培养物中检测弧菌。分子方法(PCR 和 FISH)检测所有样本均为阳性,但仅 75%的样本培养阳性,证实了这些方法在检测自然样本中的弯曲菌属的局限性。可培养的弧菌在所有时间都被回收,除了 7 月,分离出的物种仅归属于嗜冷弯曲菌。

结论

在河口环境中存在可培养和不可培养的弯曲菌属形式。PCR 检测比传统培养更能检测到弯曲杆菌和嗜冷弯曲菌。与 PCR 技术相比,FISH 可以提供关于单个细胞形态和活力的信息。

研究的意义和影响

我们的研究表明,在意大利河口地区存在潜在致病性弧菌的环境储库,它们可能以存活但不可培养的状态存在。

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