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针对金黄色葡萄球菌鉴定的不同标志物的聚合酶链反应准确性:一项以基质辅助激光解吸/电离飞行时间质谱作为金标准的比较研究

Accuracy of PCR targeting different markers for Staphylococcus aureus identification: a comparative study using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as the gold standard.

作者信息

Saraiva Mauro M, De Leon Candice M, Santos Silvana C, Stipp Danilo T, Souza Miliane M, Santos Filho Lauro, Gebreyes Wondwossen A, Oliveira Celso J

机构信息

Departments of Animal Sciences (Saraiva, De Leon, Santos, Oliveira), College of Agricultural Sciences, Federal University of Paraiba, Areia, Paraíba, Brazil.

Veterinary Sciences (Stipp), College of Agricultural Sciences, Federal University of Paraiba, Areia, Paraíba, Brazil.

出版信息

J Vet Diagn Invest. 2018 Mar;30(2):252-255. doi: 10.1177/1040638717732370. Epub 2017 Nov 6.

Abstract

Staphylococcus aureus is considered a major pathogen in veterinary and human medicine, and the emergence of multidrug-resistant strains, such as livestock-associated methicillin-resistant S. aureus, means that reliable, inexpensive, and fast methods are required to identify S. aureus obtained from animal sources. We tested the accuracy of a PCR targeting the genes femA, nuc, and coa in identifying S. aureus from animals. A total of 157 Staphylococcus spp. isolates were examined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry; 18 different Staphylococcus species were identified. Of 68 S. aureus isolates, the genes femA, nuc, and coa were found in 61, 53, and 32 isolates, respectively. Considering MALDI-TOF as the gold standard, the PCR assays targeting all 3 genes showed 100% specificity; the sensitivity values were 89.7, 77.9, and 47.0% for femA, nuc, and coa, respectively. Sensitivity was 100% when femA and nuc markers were targeted simultaneously. These results confirm PCR as an accurate method to identify S. aureus species from animal sources and strongly suggest the simultaneous use of primers targeting femA and nuc genes.

摘要

金黄色葡萄球菌被认为是兽医和人类医学中的主要病原体,而多重耐药菌株的出现,如与家畜相关的耐甲氧西林金黄色葡萄球菌,意味着需要可靠、廉价且快速的方法来鉴定从动物源获得的金黄色葡萄球菌。我们测试了针对femA、nuc和coa基因的聚合酶链反应(PCR)在鉴定动物源金黄色葡萄球菌方面的准确性。通过基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱法对总共157株葡萄球菌属分离株进行了检测;鉴定出了18种不同的葡萄球菌。在68株金黄色葡萄球菌分离株中,分别在61、53和32株分离株中发现了femA、nuc和coa基因。以MALDI-TOF作为金标准,针对所有3个基因的PCR检测显示特异性为100%;femA、nuc和coa的灵敏度值分别为89.7%、77.9%和47.0%。当同时靶向femA和nuc标记时,灵敏度为100%。这些结果证实PCR是一种从动物源鉴定金黄色葡萄球菌的准确方法,并强烈建议同时使用靶向femA和nuc基因的引物。

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