Exploration of MLVA Genotyping Application in Brucella Clinical Isolates
LYU Yan-ning1,2, CHEN Li-juan1, LI Ren-qing1, SUN Yu-lan1,CHEN Yan-wei1,WANG Quan-yi1
1. Beijing Center for Diseases Prevention and Control, Beijing Research Centre for Preventive Medicine,Beijing 100013, China; 2. School of Public Health, Capital Medical University, Beijing 100069, China
Abstract:To investigate the significance of MLVA genotyping for the discrimination of brucellosis relapse and reinfection. Brucella strains were isolated from the patients with primary and secondary onset of brucellosis. The Brucella Species identification and the MLVA-16 genotyping were carried out for the isolates, the results were then analyzed and the differences between the genotypes and each VNTR were investigated. Eight Brucella strains were isolated from four patients with double onsets of brucellosis. Seven genotypes were identified by MLVA-16 genotyping for the eight isolates. Two strains isolated from one patient with a short time interval of the double onsets was identified as the same MLVA-16 genotype, which indicated that the secondary onset was not reinfection but relapse. While the other six stains isolated from the other patients with a longer time interval of the double onsets were identified as six different MLVA-16 genotypes, and each strain was shown in a difference at one locus. The results indicated that the secondary onset of these three patients were likely reinfection but not relapse. MLVA genotyping of the isolates from the patients with double onsets of brucellosis could provide clue for the discrimination of brucellosis relapse and reinfection.
[1] 国务院. 中华人民共和国传染病防治法[J]. 中华人民共和国国务院公报,2004,31: 4-15. [2] 王大力,李晔.布鲁杆菌病防控策略的回顾与思考[J].中华地方病学杂志,2015,34(5):313-314. DOI: 10.3760/cma.j.issn.2095-4255.2015.05.001 [3] 刘锋,金福芝,江伟,等.浅析影响我国人间布鲁菌病防控的原因[J].中华地方病学杂志,2016,35(5):386-389. DOI: 10.3760/cma.j.issn.2095-4255.2016.05.018 [4] 跃华,张海洋,海岩,等.2015-2018年内蒙古自治区流行性感冒监测结果分析[J].国际病毒学杂志,2018,25(5): 292-295. DOI: 10.3760/cma.j.issn.2095-4255.2016.05.018 [5] 张艺,王晓春,张曼,等.2016年乌鲁木齐市丙型病毒性肝炎诊断及报告数据质量核查分析[J].国际病毒学杂志. 2018, 25(1):19-24. DOI: 10.3760/cma.j.issn.1673-4092.2018.01.005 [6] 郭正印,吕家锐,郝宗宇,等.71例复发性布鲁氏菌病的流行病学与临床调查报告[J].疾病预防控制通报,2013, 28(2):57-59,69. [7] 樊永贞,王德富,沈跃文,等.急、慢性布鲁杆菌病患者临床特征比较分析[J].疾病监测与控制杂志,2015, 9(11):761-762. [8] 吕燕宁,郭潇潇,陈丽娟,等.两株布鲁菌MLST新序列型(ST)的鉴定[J].中国人兽共患病学报,2018,34(5):434-440,446. DOI: 10.3969/j.issn.1002-2694.2018.00.059 [9] 姜海,荣蓉,田国忠,等. 利用多重聚合酶链反应和多色毛细管电泳分析布氏菌多位点可变数目串联重复序列[J].中国人兽共患病学报,2015,31(4):303-310. DOI: 10.3969/j.issn.1002-2694.2015.04.003 [10] Cross AR, Baldwin VM, Roy S, et al.Zoonoses under our noses[J]. Microbes Infect, 2019, 21(1): 10-19. DOI: 10.1016/j.micinf.2018.06.001 [11] Le Fleche P, Jacques I, Grayon M, et al.Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay[J]. BMC microbiology, 2006,6:9. DOI: 10.1186/1471-2180-6-9 [12] Al Dahouk S, Fleche PL, Nockler K, et al.Evaluation of Brucella MLVA typing for human brucellosis[J]. J Microbiol methods, 2007, 69(1):137-145. DOI: 10.1016/j.mimet.2006.12.015 [13] Garcia-Yoldi D, Le Fleche P, De Miguel MJ, et al.Comparison of multiple-locus variable-number tandem-repeat analysis with other PCR-based methods for typing Brucella suis isolates[J]. J Clin Microbiol, 2007, 45(12):4070-4072. DOI: 10.1128/JCM.01096-07 [14] 刘志国, 崔步云, 刘日宏, 等. 布鲁菌分子分型方法应用研究进展[J]. 中国人兽共患病学报, 2015,31(2): 163-168. DOI: 10.3969/ j.issn.1002-2694.2015.02.015 [15] Liu ZG,Wang LJ,Piao DR,et al.Molecular investigation of the transmission pattern of Brucella suis 3 from Inner Mongolia, China[J]. Front Vet Sci, 2018,5(29):271. DOI: 10.3389/fvets.2018.00271 [16] Ma JY, Wang H, Zhang XF, et al.MLVA and MLST typing of Brucella from Qinghai, China[J]. Infect Dis poverty,2016,5(1):26. DOI: 10.1186/s40249-016-0123-z [17] Jiang H,Wang H,Xu L,et al.MLVA genotyping of Brucella melitensis and Brucella abortus isolates from different animal species and humans and identification of Brucella suis vaccine strain S2 from cattle in China[J]. PLoS One,2013,8(10):e76332. DOI: 10.1371/journal.pone.0076332 [18] Xiao P, Yang H, Di D, et al.Genotyping of human Brucella melitensis biovar 3 isolated from Shanxi Province in China by MLVA16 and HOOF[J]. PLoS One,2015,10(1):e0115932. DOI: 10.1371/journal.pone.0115932 [19] Sun M J, Di DD, Li Y, et al.Genotyping of Brucella melitensis and Brucella abortus strains currently circulating in Xinjiang, China[J]. Infect Genet Evol,2016,44:522-529. DOI: 10.1016/j.meegid.2016.07.025 [20] Sun M, Jing Z, Di D, et al.Multiple locus variable-number tandem-repeat and single-nucleotide polymorphism-based Brucella typing reveals multiple lineages in Brucella melitensis currently endemic in China[J]. Front Vet Sci, 2017, 4: 215. DOI: 10.3389/fvets.2017.00215 [21] Liu ZG, Di DD, Wang M, et al.MLVA genotyping characteristics of human Brucella melitensis isolated from Ulanqab of Inner Mongolia,China[J]. Front Microbiol, 2017, 8: 6. DOI: 10.3389/fmicb.2017.00006 [22] Kilic S,Ivanov IN,Durmaz R,et al.Multiple-locus variable-number tandem-repeat analysis genotyping of human Brucella isolates from turkey[J]. J Clin Microbiol,2011,49(9):3276-3283. DOI: 10.1128/JCM.02538-10