Feasibility analysis of identification of Mycobacterium bovis by thiophen-2-carboxylic acid hydrazine biochemical test

doi:10.19982/j.issn.1000-6621.20240543

Abstract

Abstract:

Objective: To evaluate the reliability of using thiophene-2-carboxylic acid hydrazide medium for identifying Mycobacterium bovis. Methods: One thousand two hundred and sixty-seven clinically isolated mycobacterium strains collected from 14 tuberculosis prevention and control institutions and 6 designated hospitals in Beijing from January 1 to December 31, 2019 were identified by medium containing P-nitrobenzoic acid and thiophen-2-carboxylic acid hydrazide. The strains of Mycobacterium tuberculosis complex (MTBC) with fewer than 20 colonies growing on culture medium containing 5 μg/ml thiophene-2-carboxylic acid hydrazine were re-inoculated into roche culture medium containing 5 μg/ml, 2.5 μg/ml, 1.25 μg/ml and 0.63 μg/ml thiophene-2-carboxylic acid hydrazine from another reagent company for sensitivity testing. At the same time, these strains were amplified by PCR and the products were subjected to agarose gel electrophoresis to determine whether they were Mycobacterium bovis by the presence of 168 bp bands. Results: Of the 1267 clinical mycobacterium isolates, 1177 strains belonged to MTBC, and 148 of these strains formed fewer than 20 colonies after culture on the medium containing 5 μg/ml of thiophene-2-carboxylic acid hydrazine. After changing into using medium from another reagent company, there were 27 sensitive strains in the 5 μg/ml medium and 10 sensitive strains in a lower concentration (2.5 μg/ml) of the same medium. In addition, molecular biological PCR experiments showed that only 1 of the 148 isolates was identified as Mycobacterium bovis. Conclusion: The error rate of thiophene-2-carboxylic acid hydrazine medium for identifying Mycobacterium bovis was high, so this method could not be used as a reliable identification evidence.

Key words: Mycobacterium bovis, Biochemistry, Feasibility studies

CLC Number:

R378.91

Yang Xinyu, Chen Hao, Dai Xiaowei, Li Qiao, Pang Mengdi, Li Bo, Miao Honghao, Li Jie, Li Chuanyou. Feasibility analysis of identification of Mycobacterium bovis by thiophen-2-carboxylic acid hydrazine biochemical test[J]. Chinese Journal of Antituberculosis, 2025, 47(7): 901-906. doi: 10.19982/j.issn.1000-6621.20240543

Figures/Tables 3

References 11

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序号	估算对照菌落生长数	TCH浓度5μg/ml		TCH浓度2.5μg/ml		TCH浓度1.25μg/ml		TCH浓度0.63μg/ml
序号	估算对照菌落生长数	菌落数	百分比(%)^a	菌落数	百分比(%)^a	菌落数	百分比(%)^a	菌落数	百分比(%)^a
1	700	6	0.86	10	1.43	500	71.43	700	100.00
2	500	0	0.00	0	0.00	40	8.00	400	80.00
3	600	1	0.17	15	2.50	600	100.00	600	100.00
4	800	0	0.00	0	0.00	0	0.00	500	62.50
5	700	1	0.14	0	0.00	200	28.57	700	100.00
6	600	2	0.33	20	3.33	600	100.00	600	100.00
7	600	0	0.00	20	3.33	600	100.00	600	100.00
8	400	0	0.00	6	1.50	300	75.00	400	100.00
9	600	0	0.00	15	2.50	600	100.00	600	100.00
10	600	0	0.00	10	1.67	600	100.00	600	100.00
11	700	1	0.14	5	0.71	700	100.00	700	100.00
12	1000	1	0.10	3	0.30	400	40.00	1000	100.00
13	500	3	0.60	10	2.00	500	100.00	500	100.00
14	900	0	0.00	0	0.00	100	11.11	300	33.33
15	500	0	0.00	2	0.40	20	4.00	500	100.00
16	500	0	0.00	0	0.00	4	0.80	500	100.00
17	600	0	0.00	0	0.00	0	0.00	700	116.67
18	500	1	0.20	20	4.00	300	60.00	500	100.00
19	1000	4	0.40	8	0.80	800	80.00	1000	100.00
20	1000	5	0.50	100	10.00	800	80.00	800	80.00
21	700	0	0.00	30	4.29	700	100.00	700	100.00
22	900	3	0.33	20	2.22	900	100.00	900	100.00
23	600	3	0.50	20	3.33	600	100.00	600	100.00
24	700	5	0.71	10	1.43	700	100.00	700	100.00
25	700	2	0.29	20	2.86	700	100.00	700	100.00
26	800	5	0.63	30	3.75	800	100.00	800	100.00
27	500	2	0.40	30	6.00	400	80.00	500	100.00