Development of a one-step rapid immunochromatographic test kit for detecting bovine mastitis-causing Staphylococcus aureus and streptococci

IF 1.8 3区农林科学 Q1 VETERINARY SCIENCES

Research in veterinary science Pub Date : 2025-09-23 DOI:10.1016/j.rvsc.2025.105911

Yusaku Tsugami , Aoi Sugiyama , Yu Ando , Kotaro Oda , Koji Maehana , Shintarou Hayashi , Kento Fujii , Yuya Nagasawa

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引用次数: 0

Abstract

Major pathogens causing mastitis, such as Staphylococcus aureus and streptococci can be transmitted between cows. Specific antibiotics are needed to exterminate these pathogens. However, bacterial culture for pathogen identification requires at least 18 h. Thus, a rapid diagnostic method is needed for identifying pathogens. A previously reported immunochromatographic strip (ICS) test, based on a lateral flow assay, uses individual lysis buffers for different mastitis-causing pathogens, which may produce technical errors. In this study, a one-step rapid ICS test kit using simple procedures to detect bovine mastitis-causing S. aureus and streptococci was developed. In the ICS test version (ver.) 2, the new test-strip design integrated bacteriolytic enzymes, while a common lysis buffer (consisting of 3 % Triton X-100, 0.5 % sodium N-lauroylsarcosinate, and 0.15 % Zwittergent 3–12) simplified the testing procedure. In refrigerated fresh milk samples from cows with clinical mastitis (n = 77), the sensitivity, specificity, and κ statistic values (the agreement between the results of the ICS test and bacterial culture) of the ICS test ver. 2 for S. aureus were 0.864, 0.909, and 0.752, respectively, and those for streptococci were 0.778, 0.956, and 0.700, respectively, as compared to traditional bacterial culture. Thus, S. aureus and streptococci can be detected in the milk with a simple one-step procedure. Some cross-reactivity with non-aureus staphylococci was observed in ICS test ver. 2 for S. aureus. Nonetheless, the improved ICS test kit may serve as an effective detection tool for staphylococci and streptococci mastitis, supporting timely and appropriate control of mastitis on farms.

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牛乳腺炎金黄色葡萄球菌和链球菌一步快速免疫层析检测试剂盒的研制。

引起乳腺炎的主要病原体，如金黄色葡萄球菌和链球菌，可以在奶牛之间传播。需要特定的抗生素来消灭这些病原体。然而，用于病原体鉴定的细菌培养至少需要18小时。因此，需要一种快速诊断方法来鉴定病原体。先前报道的免疫层析试纸（ICS）测试，基于横向流动试验，对不同的乳腺炎病原体使用单独的裂解缓冲液，这可能会产生技术错误。在本研究中，开发了一种使用简单程序的一步快速ICS检测试剂盒，用于检测引起牛乳腺炎的金黄色葡萄球菌和链球菌。在ICS测试版本（版本）2，新的试纸设计集成了细菌分解酶，而普通的裂解缓冲液（由3% Triton X-100, 0.5% n -月桂肌酸钠和0.15% Zwittergent 3-12组成）简化了测试程序。在临床乳腺炎奶牛的冷藏鲜奶样品（n = 77）中，ICS试验的敏感性、特异性和κ统计值（ICS试验结果与细菌培养结果的一致性）均未见显著差异。与传统细菌培养相比，金黄色葡萄球菌的2分别为0.864、0.909和0.752，链球菌的2分别为0.778、0.956和0.700。因此，金黄色葡萄球菌和链球菌可以通过简单的一步程序在牛奶中检测出来。与非金黄色葡萄球菌有交叉反应。金黄色葡萄球菌为2。尽管如此，改进后的ICS检测试剂盒可作为葡萄球菌和链球菌乳腺炎的有效检测工具，支持在农场及时和适当地控制乳腺炎。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Research in veterinary science 农林科学-兽医学

CiteScore

4.40

自引率

4.20%

发文量

312

审稿时长

75 days

期刊介绍： Research in Veterinary Science is an International multi-disciplinary journal publishing original articles, reviews and short communications of a high scientific and ethical standard in all aspects of veterinary and biomedical research. The primary aim of the journal is to inform veterinary and biomedical scientists of significant advances in veterinary and related research through prompt publication and dissemination. Secondly, the journal aims to provide a general multi-disciplinary forum for discussion and debate of news and issues concerning veterinary science. Thirdly, to promote the dissemination of knowledge to a broader range of professions, globally. High quality papers on all species of animals are considered, particularly those considered to be of high scientific importance and originality, and with interdisciplinary interest. The journal encourages papers providing results that have clear implications for understanding disease pathogenesis and for the development of control measures or treatments, as well as those dealing with a comparative biomedical approach, which represents a substantial improvement to animal and human health. Studies without a robust scientific hypothesis or that are preliminary, or of weak originality, as well as negative results, are not appropriate for the journal. Furthermore, observational approaches, case studies or field reports lacking an advancement in general knowledge do not fall within the scope of the journal.