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用于快速检测鸡肉样本中沙门氏菌的改良生物发光酶免疫测定法。

Improved bioluminescent enzyme immunoassay for the rapid detection of Salmonella in chicken meat samples.

作者信息

Fukuda S, Tatsumi H, Igimi S, Yamamoto S

机构信息

Research and Development Division, Kikkoman Corporation, Chiba, Japan.

出版信息

Lett Appl Microbiol. 2005;41(5):379-84. doi: 10.1111/j.1472-765X.2005.01783.x.

Abstract

AIMS

To evaluate an improved bioluminescent enzyme immunoassay (BEIA) using biotinylated firefly luciferase for the rapid detection of Salmonella in naturally contaminated chicken meat samples.

METHODS AND RESULTS

Capture agents and lipopolysaccharide (LPS) extraction reagents for Salmonella were investigated to improve the sensitivity of the BEIA. Also, the use of Oxoid SPRINT (Simple Pre-enrichment and Rapid Isolation New Technology) as a pre-enrichment and selective medium for 26-h BEIA detection of Salmonella in chicken meat samples was examined. The use of polymyxin B as a capture agent on solid support and 3-[(3-Cholamidopropyl) dimethylammonio] propanesulfonic acid (CHAPS) for extraction of the LPS facilitated sensitive detection of Salmonella. Of 120 chicken meat samples, 25 samples were positive using the improved BEIA with the SPRINT and 25 samples were positive using the SPRINT followed by the standard isolation methods.

CONCLUSIONS

The improved BEIA, in which polymxin B was used as a capture agent and CHAPS was used for extraction of the antigen, had a sensitivity of 96.0% and a specificity of 98.9% for the detection of Salmonella in chicken meat.

SIGNIFICANCE AND IMPACT OF THE STUDY

The improved BEIA combined with the SPRINT medium for the detection of Salmonella in chicken meat samples produced comparable results to the culture methods in 26 h.

摘要

目的

评估一种改进的生物发光酶免疫测定法(BEIA),该方法使用生物素化的萤火虫荧光素酶快速检测自然污染鸡肉样品中的沙门氏菌。

方法与结果

研究了沙门氏菌的捕获剂和脂多糖(LPS)提取试剂,以提高BEIA的灵敏度。此外,还考察了使用Oxoid SPRINT(简单预富集和快速分离新技术)作为预富集和选择性培养基,用于在26小时内通过BEIA检测鸡肉样品中的沙门氏菌。使用多粘菌素B作为固相支持物上的捕获剂,以及使用3-[(3-胆酰胺丙基)二甲基铵]丙烷磺酸(CHAPS)提取LPS,有助于灵敏地检测沙门氏菌。在120份鸡肉样品中,使用改进的BEIA结合SPRINT检测出25份阳性样品,使用SPRINT然后采用标准分离方法也检测出25份阳性样品。

结论

改进的BEIA中,多粘菌素B用作捕获剂,CHAPS用于提取抗原,对鸡肉中沙门氏菌检测的灵敏度为96.0%,特异性为98.9%。

研究的意义和影响

改进的BEIA结合SPRINT培养基用于检测鸡肉样品中的沙门氏菌,在26小时内产生的结果与培养方法相当。

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