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Use of biochemical miniaturized galleries, rRNA based lateral flow assay and Real Time PCR for Cronobacter spp. confirmation

机译：生化小型画廊的使用，基于rRNA的侧向流动分析和实时PCR用于Cronobacter spp。确认

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Identification of Cronobacter represent a major challenge for laboratories testing powdered infant formula (PIF). In the present study, two biochemical galleries and three molecular methods have been applied to confirm 276 Cronobacter spp. and non-Cronobacter isolates from different sources. Using the latest database of API 20 E and ID 32 E biochemical miniaturized kits, 53% and 78% of the isolates were identified respectively. From the available results, total accuracy for Cronobacter detection was in 97% (API 20 E) and 99% (ID 32 E). The three molecular methods were based on rRNA based lateral flow, Real Time PCR combined with either a hybridization or hydrolysis probe. For all three methods total accuracy was more than 99%. A pilot trial using Next Generation Sequencing (NGS) correctly identified 58 out of 66 isolates (88%) in DNA mixtures. The results indicate that the commercially available approaches such as ID 32 E, rRNA based lateral flow and Real Time PCR are all suitable for Cronobacter identification at the genus level. The NGS method may become a suitable alternative in the future, provided that the sequence database is improved.
机译：克隆杆菌的鉴定是实验室测试婴儿配方奶粉（PIF）的主要挑战。在本研究中，两个生化画廊和三个分子方法已被应用于确认276克罗诺杆菌属。和来自不同来源的非克罗诺杆菌分离物。使用API​​ 20 E和ID 32 E生化试剂盒的最新数据库，分别鉴定出53％和78％的分离株。从可获得的结果来看，克罗诺杆菌检测的总准确度分别为97％（API 20 E）和99％（ID 32 E）。这三种分子方法基于基于rRNA的侧向流动，实时PCR与杂交或水解探针的结合。对于这三种方法，总准确度均超过99％。使用下一代测序（NGS）进行的一项先导试验正确地鉴定了DNA混合物中66个分离株中的58个（88％）。结果表明，可商购获得的方法（例如ID 32 E，基于rRNA的侧向流和实时PCR）都适合在属水平上鉴定克氏杆菌。如果改进了序列数据库，则NGS方法将来可能会成为合适的替代方法。

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