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基于胶体金纳米颗粒的免疫层析试纸条对临床乳腺炎乳汁的快速检测

Rapid Detection From Clinical Mastitis Milk by Colloidal Gold Nanoparticle-Based Immunochromatographic Strips.

作者信息

Nagasawa Yuya, Kiku Yoshio, Sugawara Kazue, Yabusaki Naohiro, Oono Kazuyoshi, Fujii Kento, Suzuki Takahide, Maehana Koji, Hayashi Tomohito

机构信息

Dairy Hygiene Unit, Division of Pathology and Pathophysiology, Hokkaido Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, Sapporo, Japan.

NOSAI Minami, Ebetsu, Japan.

出版信息

Front Vet Sci. 2020 Jan 22;6:504. doi: 10.3389/fvets.2019.00504. eCollection 2019.

Abstract

Rapid diagnostic technologies for bovine mastitis caused by () are urgently needed. In the current study, we generated an anti-ribosomal protein-L7/L12 antibody to detect and an anti-ribosomal protein-L7/L12 antibody-coated immune-chromatographic strip (ICS) test. Moreover, we determined the ability of the ICS test to detect from milk samples collected from cows with clinical mastitis. The developed ICS reacted to in a bacteria load-dependent manner with a detection limit of ~10 CFU/mL. In the evaluation of possible cross-reactivity of the ICS test, six strains of coagulase-negative Staphylococci showed slightly positive reactions, although at a lower level; however, other bacteria were completely negative. Next, we investigated the sensitivity and specificity of the ICS test compared with the bacteriological culture method using milk samples from clinical bovine mastitis. The results of the experiments demonstrated that the ICS test had high sensitivity [100%, 95% confidence interval (CI): 91.3-100%] and specificity (91.9%, CI: 90.5-91.9%) compared with culture tests. In addition, the kappa statistic demonstrated that ICS tests showed substantial agreement (k = 0.77, CI: 0.66-0.87) with culture tests. Positive correlations were observed for the statistical analysis between ( gene) copy numbers and ICS test scores in mastitic milk infected by . Therefore, we assume that this new detection method using ICS may be useful as a highly sensitive -screening method for the diagnosis of bovine mastitis. Our findings support the ongoing effort to develop an ICS method for bovine -induced mastitis, which can contribute to the rapid diagnosis of this disease.

摘要

迫切需要针对由()引起的牛乳腺炎的快速诊断技术。在当前研究中,我们制备了一种抗核糖体蛋白-L7/L12抗体来检测(),并进行了一种抗核糖体蛋白-L7/L12抗体包被的免疫层析试纸条(ICS)检测。此外,我们测定了ICS检测从临床乳腺炎奶牛采集的牛奶样本中()的能力。所开发的ICS与()以细菌载量依赖的方式发生反应,检测限约为10 CFU/mL。在评估ICS检测可能的交叉反应性时,六株凝固酶阴性葡萄球菌显示出轻微的阳性反应,尽管水平较低;然而,其他细菌则完全为阴性。接下来,我们使用临床牛乳腺炎的牛奶样本,将ICS检测与细菌培养方法进行比较,研究了ICS检测的敏感性和特异性。实验结果表明,与培养检测相比,ICS检测具有高敏感性[100%,95%置信区间(CI):91.3 - 100%]和特异性(91.9%,CI:90.5 - 91.9%)。此外,kappa统计量表明ICS检测与培养检测显示出高度一致性(k = 0.77,CI:0.66 - 0.87)。在由()感染的患乳腺炎牛奶中,(基因)拷贝数与ICS检测分数之间的统计分析观察到正相关。因此,我们认为这种使用ICS的新检测方法可能作为一种高敏感性的()筛查方法用于牛乳腺炎的诊断。我们的研究结果支持了正在进行的开发用于牛()诱导乳腺炎的ICS方法的努力,这有助于该疾病的快速诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c111/6988510/3a626511bdab/fvets-06-00504-g0001.jpg

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