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用于诊断山羊奶中布鲁氏菌病的荧光偏振测定法的评估

Evaluation of the Fluorescence Polarization Assay for the Diagnosis of Brucellosis in Goat Milk.

作者信息

Sotolongo-Rodríguez Dianelys, Gomez-Flores Ricardo, Navarro-Soto Magda Celina, Arellano-Reynoso Beatriz, Tamez-Guerra Patricia, Ramírez-Pfeiffer Carlos

机构信息

Departamento de Microbiología e Inmunología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, San Nicolás de los Garza C.P. 66450, Nuevo León, Mexico.

Departamento de Inmunología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma de Nuevo León, Escobedo C.P. 66054, Nuevo León, Mexico.

出版信息

Vet Sci. 2022 Jun 20;9(6):303. doi: 10.3390/vetsci9060303.

Abstract

The milk ring test is a detection assay for antibodies against Brucella in bovine milk. It has good sensitivity but tends to give false positive results. In this study, we standardized the application of the fluorescence polarization assay (FPA) for the detection of antibodies against B.melitensis in goat milk. We obtained negative serum and milk samples from healthy goat flocks in the northern zone of Nuevo León. Positive milk and negative, weak, and strong controls were obtained by mixing volumes of positive control serum with negative control milk. Milk samples were treated with citric acid, after which an FPA was performed. Results were then compared with the Rose Bengal test and the FPA in serum. Milk treatment allowed the quantification of antibodies in samples. Significant differences were found between the 2%, 4%, and 6% groups, compared with the control group (F3, 67 = 17.45, p < 0.0001) but not between the 2% and 4% groups (p = 0.0718). The cut-off value was 74.1 mP, with a sensitivity (Se) of 95% and a specificity (Sp) of 100%. Se and Sp values in field milk samples were 84% and 74.55%, respectively. Despite the FPA test on milk samples showed lower Se and Sp than the FPA test on serum samples, its cutoff may be adjusted. It may be recommended as a screening test in goat milk and become useful for the control and eradication of the disease.

摘要

乳环试验是一种检测牛乳中布鲁氏菌抗体的检测方法。它具有良好的敏感性,但容易出现假阳性结果。在本研究中,我们规范了荧光偏振分析(FPA)在山羊乳中检测羊种布鲁氏菌抗体的应用。我们从新莱昂州北部地区的健康山羊群中获取了阴性血清和乳样。通过将阳性对照血清与阴性对照乳混合不同体积获得阳性乳样以及阴性、弱阳性和强阳性对照。乳样用柠檬酸处理后进行FPA检测。然后将结果与血清中的玫瑰红试验和FPA进行比较。乳样处理可对样品中的抗体进行定量。与对照组相比,2%、4%和6%组之间存在显著差异(F3, 67 = 17.45,p < 0.0001),但2%组和4%组之间无显著差异(p = 0.0718)。临界值为74.1 mP,敏感性(Se)为95%,特异性(Sp)为100%。现场乳样的Se和Sp值分别为84%和74.55%。尽管对乳样进行FPA检测显示其Se和Sp低于对血清样进行FPA检测,但可以调整其临界值。它可被推荐作为山羊乳的筛查试验,对疾病的控制和根除可能有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e00/9229069/0c9ba4775394/vetsci-09-00303-g001.jpg

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