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使用SALX系统和NIHSJ - 01检测食品中的单细胞污染及最低检测限估计

Detection of Single Cell Contamination of in Foods by SALX System and NIHSJ-01 and Estimation of LOD.

作者信息

Matsuoka Hideaki, Moriyama Takatoshi, Koshimizu Natsuko, Takatani Norimasa, Yoshida Tomonori, Shimabara Yoshiomi, Hirai Tomoe, Nakajima Kazuhide, Igimi Shizunobu, Saito Mikako

机构信息

Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.

Neogen Japan K.K., 3-3-3 Minatomirai, Nishi-ku, Yokohama, Kanagawa 220-0012, Japan.

出版信息

Food Saf (Tokyo). 2025 Jun 27;13(2):23-31. doi: 10.14252/foodsafetyfscj.D-25-00008. eCollection 2025 Jun.

Abstract

A novel idea of statistical analytical procedure for the level of detection (LOD) was demonstrated by its successful application to qualitative test methods for , SALX System and NIHSJ-01. cells of a hydrogen sulfide-producing strain FSD287 and a non-hydrogen sulfide-producing strain FSD347 were added to beef and shrimp food samples using a cell sorter to achieve bacterial cell concentration of 1, 5, and 10 cfu/25g-test portion (tp). The colony forming probability (CFP) of the added cells was estimated by means of 10×10 sorting plates. All of test portions containing FSD287 or FSD347 were decided to be positive by SALX System. NIHSJ-01 using CHROMagar® (CHS) decided test portions of each of both strains to be positive, while NIHSJ-01 using desoxycholate hydrogen sulfide lactose (DHL) agar decided selectively only FSD287-test portions to be positive. All blank test portions were negative. To evaluate the level of detection at 95% probability (LOD), in addition to test results, we introduced virtual results of blank conditions approaching zero. As a result, LOD for every case was estimated to be lower than 0.326 cfu/tp indicating that both methods were able to detect 1 cfu/tp at higher than 95% probability. Therefore, our protocol for statistical analysis for LOD was feasible for the verification of the test methods that meet the requirement of detecting small number (minimum 1 cfu/25g-tp) of target micro-organisms in food test portions.

摘要

一种用于检测限(LOD)的统计分析程序的新想法,通过其成功应用于SALX系统和NIHSJ - 01的定性测试方法得到了证明。使用细胞分选仪将产硫化氢菌株FSD287和不产硫化氢菌株FSD347的细胞添加到牛肉和虾类食品样本中,以使细菌细胞浓度达到1、5和10 cfu/25g测试份(tp)。通过10×10的分选平板估算添加细胞的菌落形成概率(CFP)。SALX系统判定所有含有FSD287或FSD347的测试份为阳性。使用CHROMagar®(CHS)的NIHSJ - 01判定两种菌株的每个测试份均为阳性,而使用脱氧胆酸盐硫化氢乳糖(DHL)琼脂的NIHSJ - 01仅选择性地判定FSD287测试份为阳性。所有空白测试份均为阴性。为了评估95%概率下的检测限(LOD),除了测试结果外,我们引入了接近零的空白条件虚拟结果。结果,每种情况的LOD估计低于0.326 cfu/tp,这表明两种方法都能够以高于95%的概率检测到1 cfu/tp。因此,我们用于LOD的统计分析方案对于验证满足在食品测试份中检测少量(最低1 cfu/25g - tp)目标微生物要求的测试方法是可行的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddeb/12214205/bd4ff8140717/foodsafetyfscj-13-2-23-g001.jpg

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