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一种用于从禽肉中检测沙门氏菌的毛细管聚合酶链反应。

A capillary polymerase chain reaction for Salmonella detection from poultry meat.

作者信息

Gunaydin E, Eyigor A, Carli K T

机构信息

Department of Microbiology, Faculty of Veterinary Medicine, Uludag University, Bursa, Turkey.

出版信息

Lett Appl Microbiol. 2007 Jan;44(1):24-9. doi: 10.1111/j.1472-765X.2006.02033.x.

Abstract

AIMS

In this study, a capillary polymerase chain reaction (cPCR) was applied for Salmonella detection from poultry meat.

METHODS AND RESULTS

Salmonella detection limits of the optimized cPCR were determined with DNA templates from the samples of tetrathionate broth (TTB), Rappaport Vassiliadis broth (RVB) and selenite cystine broth (SCB) artificially contaminated with 10-fold dilutions of 6 x 10(8) CFU ml(-1) of pure Salmonella enterica ssp. enterica serovar Enteritidis 64K stock culture. Detection limits of cPCR from TTB, RVB and SCB were found as 6, 6 x 10(1) and 6 x 10(4) CFU ml(-1), respectively. In addition, detection limits of bacteriology were also determined as 6 CFU ml(-1) with TTB and SCB, and 6 x 10(1) CFU ml(-1) with RVB. A total of 200 samples, consisting of 100 chicken and 100 turkey meat samples, were tested with optimized cPCR and bacteriology. Eight and six per cent of the chicken meat samples were found to harbour Salmonella by cPCR and standard bacteriology, respectively. Of six Salmonella isolates, four belonged to serogroup D, two to serogroup B.

CONCLUSIONS

The TTB cultures of both artificially and naturally contaminated samples were found to be superior to those of RVB and SCB cultures in their cPCR results. This cPCR, utilizing template from 18-h TTB primary enrichment broth culture, takes approximately 40 min in the successful detection of Salmonella from poultry meat.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study shows that cPCR from TTB enrichment culture of poultry meat would enable rapid detection of Salmonella in laboratories with low sample throughput and limited budget.

摘要

目的

本研究应用毛细管聚合酶链反应(cPCR)检测禽肉中的沙门氏菌。

方法与结果

使用6×10⁸CFU/ml肠炎沙门氏菌肠炎血清型64K纯菌液10倍系列稀释后人工污染的四硫磺酸钠肉汤(TTB)、Rappaport Vassiliadis肉汤(RVB)和亚硒酸盐胱氨酸肉汤(SCB)样本的DNA模板,测定优化后的cPCR对沙门氏菌的检测限。发现TTB、RVB和SCB的cPCR检测限分别为6、6×10¹和6×10⁴CFU/ml。此外,还测定了细菌学检测限,TTB和SCB为6CFU/ml,RVB为6×10¹CFU/ml。对100份鸡肉和100份火鸡肉样本共200个样本进行了优化后的cPCR和细菌学检测。通过cPCR和标准细菌学方法分别检测出8%和6%的鸡肉样本含有沙门氏菌。在6株沙门氏菌分离株中,4株属于D血清群,2株属于B血清群。

结论

无论是人工污染还是自然污染样本的TTB培养物,其cPCR结果均优于RVB和SCB培养物。这种利用18小时TTB初次富集肉汤培养物模板的cPCR,在成功检测禽肉中的沙门氏菌时大约需要40分钟。

研究的意义与影响

本研究表明,对禽肉TTB富集培养物进行cPCR能够在样本通量低且预算有限的实验室中快速检测沙门氏菌。

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