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首页> 外文期刊>Journal of Invertebrate Pathology >Evaluation of different species-specific PCR protocols for the detection of Vibrio tapetis
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Evaluation of different species-specific PCR protocols for the detection of Vibrio tapetis

机译：评估不同物种特异性PCR方案检测手足弧菌

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摘要

In this study the specificity and sensitivity of three primer pairs, Jvt1-Jvt2, VtF-VtR and VtKF-VtKR, for the detection of Vibrio tapetis were evaluated in parallel using 23 V. tapetis strains isolated from different mollusc and fish species and with different geographical origin, as well as 29 representatives of related Vibrio species. The three primer pairs amplified all the V. tapetis strains, regardless of their host or geographical origin. However, with primer sets VtF-VtR and VtKF-VtKR amplification products of the expected size were obtained from chromosomal DNA of some of the non-V. tapetis bacteria tested. The sensitivity of the three PCR detection methods was also different. The detection limit obtained with primer pairs Jvt1-Jvt2 and VtF-VtR was between 1 and 10 pg DNA/PCR tube (2-20 bacterial cells per reaction). The primer set VtKF-VtKR showed a reduction of sensitivity in at least one order of magnitude. The results were highly reproducible with all primer sets when using the same thermal cycler, although some differences were observed in the results obtained in different PCR machines. Based on the findings reported here, we propose the Jvt1-Jvt2 PCR protocol as the most adequate for an accurate detection of V. tapetis in diagnostic pathology as well as in epidemiological studies of this clam pathogen
机译：在这项研究中，使用分离自不同软体动物和鱼类以及不同鱼类的23株V. tapetis菌株平行评估了三种引物对Jvt1-Jvt2，VtF-VtR和VtKF-VtKR的特异性和敏感性。地理起源，以及相关弧菌物种的29个代表。这三个引物对扩增了所有V. tapetis菌株，无论其宿主或地理来源如何。然而，使用引物组，从一些非V族的染色体DNA获得了预期大小的VtF-VtR和VtKF-VtKR扩增产物。已对tapetis细菌进行了测试。三种PCR检测方法的灵敏度也不同。用引物对Jvt1-Jvt2和VtF-VtR获得的检出限在1到10 pg DNA / PCR管之间（每个反应2-20个细菌细胞）。引物组VtKF-VtKR显示出灵敏度至少降低了一个数量级。当使用相同的热循环仪时，所有引物组的结果均具有很高的重现性，尽管在不同的PCR仪上获得的结果存在一些差异。根据此处报告的发现，我们提出Jvt1-Jvt2 PCR方案最适合在蛤path病原体的诊断病理学和流行病学研究中准确检测V. tapetis

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