实时PCR与传统培养法检测土耳其香料中大肠杆菌O157:H7的比较

Asli Ozkizilcik, M. Ateş, B. Çerçi
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摘要

本研究分析了66种不同香料样品,比较了Real-Time PCR和传统培养方法对香料中大肠杆菌O157:H7的检测效果。因此,从土耳其伊兹密尔的不同地方收集了66种不同的香料样品(原材料)。传统的培养方法包括预富集步骤、选择步骤和确认步骤。分子法分为预富集和Real-Time PCR两步。除了这些方法外,还对样品中的好氧中温细菌、大肠菌群、粪便大肠菌群进行计数,并调查沙门氏菌的存在情况,以确定样品中的总微生物负荷。结果表明,所有样品的好氧中温细菌数平均值为5.1 × 103 -2.0 × 108 CFU/g,大肠菌群数平均值为8.0 × 102 CFU/g。在所有样本中均未发现沙门氏菌。在4份样本中,Real-Time PCR检测出大肠杆菌O157:H7阳性,而培养法检测均为阴性。我们观察到Real-Time PCR的结果比传统方法更可靠。此外,Real-Time PCR法仅需20小时即可获得结果,而常规培养方法需要4天才能完成。关键词:培养法,大肠杆菌O157:H7,实时PCR,香料
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Comparison of real-time PCR and conventional cultural methods to detect Escherichia coli O157:H7 in spices in Turkey
In this study, 66 different spice samples were analyzed in order to compare Real-Time PCR and conventional cultural methods for the detection of E. coli O157:H7 in spices. Thus, 66 different spice samples (raw materials) were collected from various places in Izmir, Turkey. Conventional cultural methods included pre-enrichment step, selection step and confirmation step. The molecular method included two steps: pre-enrichment and Real-Time PCR step. Beside these methods, total aerobic mesophilic bacteria, coliform, fecal coliform in samples, were also counted and the presence of Salmonella sp. was investigated in order to determine total microbial load in samples. The results showed that aerobic mesophilic bacteria counts were 5.1x103 -2.0x108 CFU/g, coliform counts were 8.0 x 102 CFU/g for the average values of all samples. Salmonella sp. was not found in any of the samples.In four samples E. coli O157:H7 positive by Real-Time PCR were found while all of the samples were negative by cultural methods. We observed that the results of Real-Time PCR were more reliable than conventional methods. Furthermore, the results were obtained in only 20 hours by Real-Time PCR method whereas conventional cultural method was completed in 4 days. Keywords: Cultural method, E. coli O157:H7, real-time PCR, spice
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