Genotyping and identification of 30 Mycobacterium bovine isolates from cattle
DENG Ke-xin1,2, WANG Xiao-ping3, SONG Fu-yang1,2, YAN Na1,2, ZHANG Xu1,2, DENG Guang-cun1,2
1. Ningxia University, Yinchuan 750021, China; 2. Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western China, Yinchuan 750021, China; 3. The Fourth People's Hospital of Ningxia Hui Autonomous Region, Yinchuan 750021, China;
Abstract：Genotyping was used to identify 30 isolates of Mycobacterium bovine isolated from three provinces in Northwest China. The throat and nose swabs of tuberculin (PPD) positive cows were collected from large-scale dairy farms in three provinces of northwest China. After isolation and culture, 16S rRNA, MPT64 and multi-site PCR were used for typing and identification. The genotypes were analyzed by using variable-number tandem repeats analysis and the combination of Mycobacterial interspersed repetitive unit. The results showed that all the 30 bovine clinical isolates belonged to Mycobacterium genus, of which 12 belonged to Mycobacterium tuberculosis complex group and 18 belonged to non-Mycobacterium tuberculosis complex group. Among the 12 strains of Mycobacterium tuberculosis, 10 strains were Mycobacterium bovis and 2 strains were Mycobacterium. Results of MIRU-VNTR analysis showed that 12 strains of Mycobacterium tuberculosis showed 10 VNTR genotypes, and 4 strains clustered into 2 gene clusters. All the 30 isolates belonged to Mycobacterium genus, and Mycobacterium bovis was the main strain of Mycobacterium tuberculosis.
邓可新, 王晓平, 宋孚洋, 严娜, 张旭, 邓光存. 30株牛源分枝杆菌分离株基因分型鉴定[J]. 中国人兽共患病学报, 2022, 38(3): 196-202.
DENG Ke-xin, WANG Xiao-ping, SONG Fu-yang, YAN Na, ZHANG Xu, DENG Guang-cun. Genotyping and identification of 30 Mycobacterium bovine isolates from cattle. Chinese Journal of Zoonoses, 2022, 38(3): 196-202.
[1] Olea-Popelka F, Muwonge A, Perera A, et al.Zoonotic tuberculosis in human beings caused by Mycobacterium bovis-a call for action[J]. Lancet Infect Dis,2017,17(1):e21-e25. DOI:10.1016/S1473-3099(16)30139-6 [2] 许芳. 我国部分地区牛结核病调查与牛肺外结核病病原分离鉴定及生物学特性研究[D].兰州:甘肃农业大学, 2020. [3] Romha G, Gebru G, Asefa A, et al.Epidemiology of Mycobacterium bovis and Mycobacterium tuberculosis in animals: Transmission dynamics and control challenges of zoonotic TB in Ethiopia[J]. Prev Vet Med,2018,158: 1-17. DOI:10.1016/j.prevetmed.2018.06.012 [4] Khan MK, Islam MN, Ferdous J, et al.An overview on epidemiology of tuberculosis[J]. Mymensingh Med, 2019, 28(1):259-266. [5] Guimaraes AMS, Zimpel CK.Mycobacterium bovis: from genotyping to genome sequencing[J]. Microorganisms, 2020, 8(5):667. DOI:10.3390/microorganisms8050667 [6] Teeter LD, Vempaty P, Nguyen DT, et al.Validation of genotype cluster investigations for Mycobacterium tuberculosis: application results for 44 clusters from four heterogeneous United States jurisdictions[J]. BMC Infect Dis, 2016, 16(1):594. DOI:10.1186/s12879-016-1937-9 [7] Haddad N, Ostyn A, Karoui C, et al.Spoligotype diversity of Mycobacterium bovis strains isolated in France from 1979 to 2000[J]. Clin Microbiol, 2001, 39(10):3623-3632. DOI:10.1128/JCM.39.10.3623-3632.2001 [8] Haddad N, Masselot M, Durand B.Molecular differentiation of Mycobacterium bovis isolates. Review of main techniques and applications[J]. Res Vet Sci, 2004, 76(1):1-18. DOI:10.1016/s0034-5288(03)00078-x [9] 曹瑞,张喜悦,孙照刚,等.新疆牛分枝杆菌MIRU-VNTR基因分型的初步研究[J].中国预防兽医学报,2010,32(7):524-528. [10] Pignone M, Greth KM, Cooper J, et al.Identification of mycobacteria by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry[J]. J Clin Microbiol, 2006, 44(6): 1963-70. DOI:10.1128/JCM.01959-05 [11] Cloud JL, Neal H, Rosenberry R, et al.Identification of Mycobacterium spp by using a commercial 16S ribosomal DNA sequencing kit and additional sequencing libraries[J]. J Clin Microbiol, 2002, 40(2): 400-406. DOI:10.1128/JCM.40.2.400-406.2002 [12] Huard RC, Lazzarini LC, Butler WR, et al.PCR-based method to differentiate the subspecies of the Mycobacterium tuberculosis complex on the basis of genomic deletions[J]. Clin Microbiol, 2003, 41(4):1637-1650. DOI:10.1128/JCM.41.4.1637-1650.2003 [13] Vargas AN, Suazo FM, Alarcon JC, et al.Genetic diversity based on MIRU-VNTR profile of isolates of Mycobacterium bovis from Mexican cattle[J]. Prev Vet Med, 2016, 131:75-78. DOI:10.1016/j.prevetmed.2016.07.007 [14] Karahan Z C, Akar N.Restriction endonuclease analysis as a solution for determining rifampin resistance mutations by automated DNA sequencing in heteroresistant Mycobacterium tuberculosis strains[J]. Microb Drug Res, 2005, 11(2): 137. DOI:10.1089/mdr.2005.11.137 [15] World Health Organization.WHO consolidated guidelines on tuberculosis: module 1: prevention: tuberculosis preventive treatment [R]. Geneva: World Health Organization, 2020. [16] 王岩. 华北某地区牛结核病流行病学调查研究[D].南京:南京农业大学,2016. [17] 林康,田莉莉,常塔娜,等.新疆某牛场牛结核病的综合诊断与病原分离鉴定[J].中国人兽共患病学报,2019,35(12):1150-1153. DOI:10.3969/j.issn.1002-2694.2019.00.177
2022, Vol. 38 
