Christian Doppler Laboratory for Molecular Food Analytics, Veterinärplatz 1, Vienna, Austria.
Lett Appl Microbiol. 2010 Oct;51(4):480-4. doi: 10.1111/j.1472-765x.2010.02925.x.
A rapid real-time PCR-based method for the detection of Listeria monocytogenes was applied to the examination of 44 Quargel cheese samples from a recent outbreak in Austria and compared to the standard method according to ISO-16140.
The combined enrichment/real-time PCR method amplifying the prfA locus was performed according to [Rossmanith et al.(2006) Res Microbiol, 157, 763-771]. Qualitative and quantitative examination of the samples was performed according to the standard method ISO-11290. Comparison of the combined enrichment/real-time PCR method with ISO-11290 resulted in 100% relative accuracy, 100% relative sensitivity and 100% relative specificity.
A previously published study describing the validation of the method, including samples after storage at -80 degrees C, resulted in lower performance values. In contrast, the samples were stored at +4 degrees C in this study. The results of this study indicate an effect of storage, thus masking the true performance of the method.
The results of this study are discussed together with the previously published data to demonstrate the excellent qualities of this rapid (< or = 30 h) method when applied to fresh specimens stored at +4 degrees C.
应用基于实时聚合酶链反应（PCR）的快速检测李斯特菌方法，对奥地利近期暴发疫情中的 44 份夸格尔奶酪样本进行检测，并与 ISO-16140 标准方法进行比较。
采用扩增 prfA 基因座的组合增菌/实时 PCR 方法，按照[Rossmanith 等人（2006）Res Microbiol，157，763-771]进行。根据标准方法 ISO-11290 对样品进行定性和定量检测。组合增菌/实时 PCR 方法与 ISO-11290 的比较结果为相对准确度 100%、相对灵敏度 100%和相对特异性 100%。
之前发表的研究描述了该方法的验证，包括在-80°C 下储存后的样本，结果显示性能值较低。相比之下，本研究中样本在+4°C 下储存。本研究的结果表明存在储存效应，从而掩盖了该方法的真实性能。
本研究结果与之前发表的数据一起讨论，以证明该快速（≤30 小时）方法在应用于+4°C 新鲜样本时具有优异的特性。
