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首页> 外文期刊>Revista Brasileira de Ciência Avícola >Duplex Real-Time PCR Using Sybr Green I for Quantification and Differential Diagnosis between Salmonella Enteritidis and Salmonella Typhimurium
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Duplex Real-Time PCR Using Sybr Green I for Quantification and Differential Diagnosis between Salmonella Enteritidis and Salmonella Typhimurium

机译：使用Sybr Green I进行双重实时PCR定量和肠炎沙门氏菌和鼠伤寒沙门氏菌之间的鉴别诊断

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摘要

The incidence of foodborne diseases caused by the genus Salmonella spp. in industrialized countries is often high in epidemiological surveys. Obtaining a rapid diagnostic test for identification of bacteria is crucial in order to rapidly implement control measures to contain bacterial spread, to reduce losses in animal production and to avoid risks from food-borne infections to human health. The aim of this study was to standardize duplex real-time PCR using SYBr Green I for differential and quantitative diagnosis of S . Typhimurium and S . Enteritidis. According to the experiment, the melting temperature of 85°C was observed for a 206bp amplified product when S . Enteritidis DNA was added to the reaction. S . Typhimurium DNA showed that the melting temperature of 79°C when observed for a 62bp amplified product. The standard curve showed the high sensitivity of the proposed test, since it was possible to obtain eight quantification points, starting at 10 8 CFU/mL and ending at 10 1 CFU/mL. As a result of the present study, a real-time PCR duplex reaction with high sensitivity, specificity and based on the fluorescence of SYBr Green I was standardized. In addition, this methodology aligns low cost to the faster diagnostic result, in relation to other molecular tests, making it attractive for application in routine laboratory analyzes.
机译：沙门氏菌属引起的食源性疾病发生率。在工业化国家，流行病学调查通常很高。为了快速实施控制措施以遏制细菌传播，减少动物生产的损失并避免食源性感染对人类健康的风险，获取细菌的快速诊断测试至关重要。这项研究的目的是使用SYBr Green I对S的鉴别和定量诊断进行双重实时PCR标准化。鼠伤寒和S。肠炎。根据该实验，观察到当S为206bp时扩增产物的熔化温度为85℃。将肠炎沙门氏菌DNA添加到反应中。 。鼠伤寒DNA显示，观察到62bp的扩增产物的融解温度为79℃。标准曲线显示了拟议测试的高灵敏度，因为有可能获得八个定量点，起始点为10 8 CFU / mL，终点为10 1 CFU / mL。作为本研究的结果，标准化了具有高灵敏度，特异性并基于SYBr Green I荧光的实时PCR双链反应。此外，相对于其他分子测试，这种方法使低成本与更快的诊断结果保持一致，使其在常规实验室分析中具有吸引力。

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