牛分枝杆菌脂多糖在动物结核病血清学诊断中的应用价值

doi:10.19982/j.issn.1000-6621.20220496

摘要/Abstract

摘要：

目的: 评价牛分枝杆菌脂多糖在动物结核病血清学诊断中的临床应用价值。方法: 从灭活的牛分枝杆菌中提取具有生物活性的脂多糖,用提取的脂多糖进行酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA),检测阴性牛血清和结核病、副结核病、布鲁氏菌病、口蹄疫、衣原体病和新孢子虫病阳性牛血清,判断脂多糖用于动物结核病诊断的特异性。同时,收集牛血清样本245份[结核菌素皮肤试验(tuberculin skin test,TST)阳性177份,TST阴性68份],梅花鹿血清样本29份(TST阳性10份,TST阴性19份),獐子血清样本14份(TST阳性14份),通过ELISA方法检测血清中的抗体水平,以TST结果为参照,评估脂多糖在动物结核病血清学诊断中的临床应用价值。结果: 除结核病阳性牛血清外,其余阳性病牛血清的检测结果与阴性牛血清差异无统计学意义(P>0.05),且均低于Cut off值。基于脂多糖建立的ELISA抗体检测方法与TST结果的总符合率为88.54%(255/288),总敏感度为87.56%(176/201),总特异度为90.80%(79/87)。结论: 牛分枝杆菌脂多糖可作为动物结核病血清学诊断的特异性抗原之一,有助于提高现有诊断方法的特异性和检出率,具有较好的应用价值。

关键词: 分枝杆菌,牛, 脂多糖类, 结核, 血清学试验

Abstract:

Objective: To evaluate the clinical application of Mycobacterium bovis (M.bovis) lipopolysaccharide in the serological diagnosis of tuberculosis in animals. Methods: Bioactive lipopolysaccharides were extracted from inactivated M.bovis, and tested using enzyme linked immunosorbent assay (ELISA) to detect negative bovine sera and positive bovine sera for tuberculosis, paratuberculosis, brucellosis, foot-and-mouth disease, chlamydiosis and neosporosis, to determine the specificity of lipopolysaccharides for the diagnosis of animal tuberculosis. Meanwhile, 245 bovine serum samples (177 were positive and 68 were negative for tuberculin skin test (TST)), 29 plum deer serum samples (10 were positive and 19 were negative for TST) and 14 roe deer serum samples (all positive for TST) were collected. The antibody levels were measured by ELISA, and the TST results were used as a reference to evaluate the clinical application of lipopolysaccharide in the serological diagnosis of tuberculosis in animals. Results: Except for the tuberculosis positive bovine sera, the results of the positive diseased bovine sera were not statistically different from those of the negative bovine sera (P>0.05) and were lower than the Cut off values. The overall compliance of the ELISA antibody assay based on lipopolysaccharide with the TST results was 88.54% (255/288), the overall sensitivity was 87.56% (176/201), and the overall specificity was 90.80% (79/87). Conclusion: M.bovis lipopolysaccharide can be used as one of the specific antigens for the serological diagnosis of animal tuberculosis, it helps to improve the specificity and detection rate of existing diagnostic methods, and has good application value.

Key words: Mycobacterium bovis, Lipopolysaccharides, Tuberculosis, Serologic tests

中图分类号:

R52
R522.2

王元智, 龙美贞, 刘一朵, 周向梅. 牛分枝杆菌脂多糖在动物结核病血清学诊断中的应用价值[J]. 中国防痨杂志, 2023, 45(4): 401-405. doi: 10.19982/j.issn.1000-6621.20220496

Wang Yuanzhi, Long Meizhen, Liu Yiduo, Zhou Xiangmei. Application value of Mycobacterium bovis lipopolysaccharide in the serological diagnosis of tuberculosis in animals[J]. Chinese Journal of Antituberculosis, 2023, 45(4): 401-405. doi: 10.19982/j.issn.1000-6621.20220496

图/表 3

表1

表2

图1

参考文献 12

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脂多糖包被浓度	血清稀释比例
脂多糖包被浓度	不稀释	1∶10	1∶20	1∶40
10.00μg/ml
阳性血清A₄₅₀值(P)	0.236	0.493	0.488	0.366
阴性血清A₄₅₀值(N)	0.140	0.142	0.124	0.119
P/N值	1.690	3.470	3.940	3.080
5.00μg/ml
阳性血清A₄₅₀值(P)	0.196	0.276	0.333	0.431
阴性血清A₄₅₀值(N)	0.149	0.132	0.124	0.193
P/N值	1.320	2.090	2.690	2.230
2.50μg/ml
阳性血清A₄₅₀值(P)	0.235	0.257	0.354	0.306
阴性血清A₄₅₀值(N)	0.136	0.118	0.122	0.134
P/N值	1.730	2.180	2.900	2.280
1.25μg/ml
阳性血清A₄₅₀值(P)	0.305	0.186	0.130	0.129
阴性血清A₄₅₀值(N)	0.112	0.109	0.114	0.124
P/N值	2.720	1.710	1.140	1.040

检测指标	阴性	结核病	副结核病	布鲁氏菌病	口蹄疫	衣原体病	新孢子虫病
A₄₅₀值	0.178	0.699	0.185	0.175	0.182	0.147	0.173
	0.175	0.725	0.164	0.169	0.191	0.196	0.192
	0.183	0.929	0.181	0.171	0.175	0.177	0.163
	0.153	0.963	0.185	0.177	0.184	0.160	0.176
	0.174	0.849	0.172	0.161	0.165	0.187	0.177
	0.204	0.918	0.161	0.189	0.170	0.186	0.172
平均值	0.178	0.847	0.175	0.174	0.178	0.176	0.176
标准差	0.015	0.102	0.010	0.009	0.009	0.017	0.009
P值		<0.001	0.748	0.641	0.930	0.868	0.824

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