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基于重复序列的 PCR 对澳大利亚家禽源沙门氏菌血清型的分类。

Classification of Salmonella enterica serotypes from Australian poultry using repetitive sequence-based PCR.

机构信息

Birling Avian Laboratories, Bringelly, NSW, Australia.

出版信息

J Appl Microbiol. 2012 Jan;112(1):185-96. doi: 10.1111/j.1365-2672.2011.05172.x. Epub 2011 Nov 1.

DOI:10.1111/j.1365-2672.2011.05172.x
PMID:21981579
Abstract

AIMS

To evaluate a semi-automated repetitive extragenic palindromic sequence-based PCR (rep-PCR) system for the classification of Salmonella serotypes from Australian poultry.

METHODS AND RESULTS

Using a DNA fingerprint library within the DiversiLab(®) System, four separate databases were constructed (serogroup B, C, E and Other). These databases contained 483 serologically confirmed (reference laboratory) Salmonella isolates. A blinded set of Salmonella cultures (n = 155) were typed by rep-PCR, matched against the internal library and compared with traditional serotyping. The predicted (Kullback-Leibler) serotype of 143 (92·3%) isolates matched traditional typing (P < 0·05). Of the 12 (7·7%) remaining isolates, ten (6·5%) resulted in 'No Match', one (0·65%) was incorrectly matched to the library (Salm. subsp 1 ser 4,12:-:-), and the other (0·65%) was referenced as Salm. ser. Sofia, whereas rep-PCR and in-house serotyping concurred as Salmonella serovar Typhimurium. Financial analysis showed higher material cost (215%) and a lower labour component (47·5%) for rep-PCR compared with serotyping.

CONCLUSION

The DiversiLab(®) System, with serogroup databases, was successfully implemented as an adjunct for reference serotyping of Salmonella enterica.

SIGNIFICANCE AND IMPACT OF THE STUDY

The DiversiLab(®) System platform is a cost-effective and easy-to-use system, which can putatively determine Salmonella enterica serotypes within a few hours.

摘要

目的

评估一种半自动化重复外回文序列基聚合酶链反应(rep-PCR)系统,用于对澳大利亚家禽中的沙门氏菌血清型进行分类。

方法与结果

使用 DiversiLab(®)系统中的 DNA 指纹图谱库,构建了四个独立的数据库(血清群 B、C、E 和其他)。这些数据库包含了 483 个经血清学确认(参考实验室)的沙门氏菌分离株。通过 rep-PCR 对一组盲测的沙门氏菌培养物(n = 155)进行分型,与内部库进行匹配,并与传统血清分型进行比较。143 株(92.3%)分离株的预测(Kullback-Leibler)血清型与传统分型相匹配(P < 0.05)。在其余的 12 株(7.7%)分离株中,有 10 株(6.5%)结果为“无匹配”,1 株(0.65%)被错误地匹配到库中(Salm. subsp 1 ser 4,12:-:-),另 1 株(0.65%)被标注为 Salm. ser. Sofia,而 rep-PCR 和内部血清分型均确认为沙门氏菌 Typhimurium 血清型。财务分析表明,与血清分型相比,rep-PCR 的材料成本更高(215%),劳动力成分更低(47.5%)。

结论

血清群数据库的 DiversiLab(®)系统成功地作为沙门氏菌参考血清分型的辅助手段得以实施。

研究的意义和影响

DiversiLab(®)系统平台是一种具有成本效益且易于使用的系统,可在数小时内推测出沙门氏菌血清型。

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