Tominaga Tatsuya
Saitama Industrial Technology Center North Institute, 2-133, Suehiro, Kumagayashi, Saitama 360-0031, Japan.
J Microbiol Methods. 2018 Apr;147:43-49. doi: 10.1016/j.mimet.2018.02.015. Epub 2018 Mar 6.
Bacteria of the genera Klebsiella and Raoultella, which are present in foods and the natural environment, are associated with health hazards in humans. In the present study, two types of strips-based methods were developed to detect these bacteria simply and quickly. One method used lateral-flow test strips (LFTS) in combination with anti-Klebsiella antibodies labeled with palladium nanoparticles that bind to target bacteria, allowing their visualization. In the other, the antibodies were immobilized on nitrocellulose membranes, and urease activity was measured using an ion sensitive field effect transistor pH sensor. Testing of a combination of these two methods on 72 cultured strains successfully identified all 25 strains of Klebsiella pneumoniae, Klebsiella oxytoca and Raoultella ornithinolytica. This approach also accurately identified 76 of 77 (99%) strains isolated from meats and pastries. When combined with preculture, this method was accurate in identifying 19 of 26 (73%) target bacteria in food. These results suggest that the novel combination of strip-based assays may be effective for the on-site monitoring of food production plants, and thereby enhance food safety.
克雷伯氏菌属和拉乌尔菌属细菌存在于食品和自然环境中，与人类健康危害相关。在本研究中，开发了两种基于试纸条的方法来简单快速地检测这些细菌。一种方法是将侧向流动试纸条（LFTS）与用钯纳米颗粒标记的抗克雷伯氏菌抗体结合使用，该抗体与目标细菌结合，使其可视化。另一种方法是将抗体固定在硝酸纤维素膜上，并使用离子敏感场效应晶体管pH传感器测量脲酶活性。对这两种方法组合对72株培养菌株进行测试，成功鉴定出所有25株肺炎克雷伯菌、产酸克雷伯菌和解鸟氨酸拉乌尔菌。该方法还准确鉴定出从肉类和糕点中分离出的77株菌株中的76株（99%）。与预培养相结合时，该方法在鉴定食品中26株目标细菌中的19株（73%）时是准确的。这些结果表明，基于试纸条检测的新组合可能对食品生产工厂的现场监测有效，从而提高食品安全。
