COMPARISON OF METHODS FOR PROCESSING LITTER AND AIR FILTER MATRICES FROM POULTRY HOUSES TO OPTIMIZE POLYMERASE CHAIN REACTION DETECTION OF SALMONELLA TYPHIMURIUM

Y.M. KWON, C.L. WOODWARD, J. PEÑA, D.E. CORRIER, S.D. PILLAI, S.C. RICKE
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引用次数: 11

Abstract

Abstract An integrated effort to reduce the occurrence of Salmonella in poultry products should include the control of Salmonella in poultry production in early stages, which requires an efficient monitoring of Salmonella spp. in poultry houses. Studies were conducted to evaluate the potential advantages of using air samples as an alternative to litter samples for PCR detection of Salmonella spp. from poultry houses. The air filter and litter samples from a poultry house were spiked with a culture of Salmonella typhimurium. The intact samples and the DNA extracted from the samples were used for amplification of Salmonella-specific DNA using hns gene specific primers. The ***hns gene was more effectively amplified with filter samples than litter samples when the intact sample was used as template. When extracted DNA was used, however, there was no difference between the two samples in their intensities of PCR products as visualized on an agarose gel. The dilution of the matrices from both sampling methods greatly increased the amplification efficiency, indicating that it was the background matrices that hindered the PCR amplification. The results in this study suggest that air filter sampling may be a labor-saving alternative to litter sampling, which can be used to increase the sensitivity of PCR detection of Salmonella spp. without prior sample processing.

鸡舍垃圾和空气过滤基质处理方法的比较优化鼠伤寒沙门菌聚合酶链反应检测
减少家禽产品中沙门氏菌发生的综合努力应包括在家禽生产的早期阶段控制沙门氏菌,这需要对家禽舍中的沙门氏菌进行有效监测。进行了研究,以评估使用空气样本作为替代垃圾样本的潜在优势,用于PCR检测家禽舍沙门氏菌。从一个家禽舍的空气过滤器和垃圾样本中加入了鼠伤寒沙门氏菌培养物。完整的样品和提取的DNA使用hns基因特异性引物扩增沙门氏菌特异性DNA。以完整样品为模板,过滤样品比凋落物样品更有效地扩增***hns基因。然而,当使用提取的DNA时,在琼脂糖凝胶上观察到,两种样品的PCR产物强度没有差异。两种取样方法对基质的稀释度都大大提高了扩增效率,表明是背景基质阻碍了PCR扩增。本研究结果表明,空气过滤器取样可能是一种省力的选择,可用于提高沙门氏菌PCR检测的灵敏度,而无需事先处理样品。
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来源期刊
Journal of Rapid Methods and Automation in Microbiology
Journal of Rapid Methods and Automation in Microbiology 生物-生物工程与应用微生物
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