114例HIV/MTB双重感染病例结核分枝杆菌Spoligotyping基因分型研究
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摘要
目的了解HIV/MTB双重感染病例中结核分枝杆菌基因型构成情况以及北京家族基因型菌株所占的比例,为HIV/MTB双重感染病例结核病预防控制提供分子流行病学依据。方法收集HIV/MTB双重感染病例结核分枝杆菌临床分离株114株,利用间隔寡核苷酸分型(Spoligotyping)技术对所收集的临床分离株进行分型研究,分析HIV/MTB双重感染病例中结核分枝杆菌基因型构成情况及主要流行型。结果 114例HIV/MTB双重感染病例结核分枝杆菌成2个大的基因家族,即北京家族(Beijing Family)和非北京家族(non-Beijing Family),分别占66.7%(76/114)和33.3%(38/114),共30种基因型。其中,非北京家族包括T家族共18株,H家族共5株,EAI5基因家族1株,CAS1_DEHLI基因家族1株及12种在数据库中没有对应基因家族或没有匹配的结果的新基因型13株。结论 HIV/MTB双重感染病例中,结核分枝杆菌基因型呈明显的多态性,北京家族基因型菌株仍为HIV/MTB双重感染病例中结核分枝杆菌主要流行菌株。
In the study,we aimed to understand the genotype of Mycobacterium tuberculosis in HIV/MTB co-infection patients and the proportion of Beijing family genotypes,and provide the molecular epidemiological basis for tuberculosis prevention and control in HIV/MTB cases.A total of 114 clinical isolates of Mycobacterium tuberculosis were collected from HIV/MTB patients,and then the spoligotyping technique was used to classify the collected clinical isolates and analyze the genotype composition and main epidemic strains of Mycobacterium tuberculosis in those clinical cases.Results showed the 114 clinical isolates were divided into two large gene families,the Beijing family and non-Beijing family,accounting for 66.7%(76/114)and 33.3%(38/114),respectively,and a total of 30 genotypes.Among them,the non-Beijing family included 18 strains of T family,5strains of H family,1strain of EAI5 family,1strain of CAS1_DEHLI family and 13 strains of 12 new genotypes without corresponding gene family or no matching result in the database.Our study suggests that the HIV/MTB co-infection cases showed obvious polymorphisms in the Mycobacterium tuberculosis genotype,and the Beijing family genotype strain is still the major epidemic strains.
In the study,we aimed to understand the genotype of Mycobacterium tuberculosis in HIV/MTB co-infection patients and the proportion of Beijing family genotypes,and provide the molecular epidemiological basis for tuberculosis prevention and control in HIV/MTB cases.A total of 114 clinical isolates of Mycobacterium tuberculosis were collected from HIV/MTB patients,and then the spoligotyping technique was used to classify the collected clinical isolates and analyze the genotype composition and main epidemic strains of Mycobacterium tuberculosis in those clinical cases.Results showed the 114 clinical isolates were divided into two large gene families,the Beijing family and non-Beijing family,accounting for 66.7%(76/114)and 33.3%(38/114),respectively,and a total of 30 genotypes.Among them,the non-Beijing family included 18 strains of T family,5strains of H family,1strain of EAI5 family,1strain of CAS1_DEHLI family and 13 strains of 12 new genotypes without corresponding gene family or no matching result in the database.Our study suggests that the HIV/MTB co-infection cases showed obvious polymorphisms in the Mycobacterium tuberculosis genotype,and the Beijing family genotype strain is still the major epidemic strains.
引文
[1]WHO.Global Tuberculosis Report 2016[R].WHO,2016.DOI:apps. who. int/iris/bitstream/handle/10665/250441/9789241565394-eng. pdf; jsessionid=FF724D6C10764-B149465961FB9DBF189
[2]王晓萌,刘志广,柳正卫,等.70株浙江省结核分枝杆菌临床分离株Spoligotyping基因分型研究[J].中国预防医学杂志,2008,9(11):946-949.DOI:10.3969/j.issn.1009-6639.2008.11.003
[3]王晓萌,刘志广,柳正卫,等.Spoligotyping和MLVA用于71株浙江省结核分枝杆菌临床分离株基因分型的初步研究[J].中国人兽共患病学报,2008,24(12):1090-1094.DOI:10.3969/j.issn.1002-2694.2008.12.002
[4]Kamerbeek J,Schouls L,Kolk A,et al.Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology[J].J Clin Microbiol,1997,35:907-914.
[5]Van Soolingen D.Molecular epidemiology of tuberculosis and other mycobacterial infections:main methodologies and achievements[J].J Internal Med,2001,249:1-26.DOI:10.1046/j.1365-2796.2001.00772.x
[6]Sola C,Filliol I,Legrand E,et al.Genotyping of the Mycobacterium tuberculosis complex using MIRUs:association with VNTR and spoligotyping for molecular epidemiology and evolutionary genetics[J].Infect Genet Evol,2003,3:125-133.DOI:10.1016/s1567-1348(03)00011-x
[7]杨修军,万康林,王艳华,等.应用Spoligotyping技术对吉林省结核分枝杆菌基因分型的研究[J].中国卫生工程学,2008,7(6):326-328.DOI:10.3969/j.issn.1671-4199.2008.06.002
[8]刘飞鹰,刘志广,王喜文,等.Spoligotyping对广西地区208株结核分枝杆菌临床分离株的基因分型[J].中国人兽共患病学报,2007,23(12):1226-1230.DOI:10.3969/j.issn.1002-2694.2007.12.015
[9]刘洁,同重湘,李君莲,等.中国西北部两省区结核分枝杆菌临床分离株Spoligotyping分型初步分析[J].实用预防医学,2014,21(5):522-525.DOI:10.3969/j.issn.1006-3110.2014.05.004
[10]Liu J,Tong CX,Liu J,et al.First insight into the genotypic diversity of clinical Mycobacterium tuberculosis isolates from Gansu province,China[J].PLoS ONE,2014,9(6):e99357.DOI:10.1371/journal.pone.0099357
[11]杨筠,董海燕,李定越,等.四川省106株结核分枝杆菌spoligotyping基因分型研究[J].医学研究杂志,2012,41(5):28-31.DOI:10.3969/j.issn.1673-548X.2012.05.010
[12]Jiao WW,Mokrousov I,Sun GZ,et al.Evaluation of new variable-number tandem-repeat systems for typing Mycobacterium tuberculosis with Beijing genotype isolates from Beijing,China[J].J Clin Microbiol,2008,46(3):1045-1049.DOI:10.1128/jcm.01869-07
[13]Wang J,Liu Y,Zhang CL,et al.Genotypes and characteristics of clustering and drug susceptibility of Mycobacterium tuberculosis isolates collected in Heilongjiang province,China[J].J Clin Microbiol,2011,49(4):1354-1362.DOI:10.1128/jcm.02274-10
[14]Liu Q,Yang DD,Xu WG,et al.Molecular typing of mycobacterium tuberculosis isolates circulating in Jiangsu Province,China[J].BMC Infectious Diseases,2011,11:288.DOI:10.1186/1471-2334-11-288
[15]Lu W,Lv B,Liu Q,et al.Genotypes of Mycobacterium tuberculosis isolates in rural China:Using MIRU-VNTR and spoligotyping methods[J].Scand J Infect Dis,2014,46:98-106.DOI:10.3109/00365548.2013.858182
[16]Yu Q,Su YK,Lu B,et al.Genetic diversity of Mycobacterium tuberculosis isolates from inner mongolia,China[J].PLoS ONE,2013,8(5):e57660.DOI:10.1371/journal.pone.0057660
[17]Ma X,Wang HY,Deng YF,et al.rpoB gene mutations and molecular characterization of rifampin-resistant Mycobacterium tuberculosis isolates from Shandong province,China[J].J Clin Microbiol,2006,44(9):3409-3412.DOI:10.1128/jcm.00515-06
[18]Han H,Wang F,Xiao Y,et al.Utility of mycobacterial interspersed repetitive unit typing for differentiating Mycobacterium tuberculosis isolates in Wuhan,China[J].J Med Microbiol,2007,56,1219-1223.DOI:10.1099/jmm.0.47005-0
[19]Dong H,Shi L,Zhao X,et al.Genetic Diversity of Mycobacterium tuberculosis Isolates from Tibetans in Tibet,China[J].PLoS ONE,2012,7(3):e33904.DOI:10.1371/journal.pone.0033904
[20]Kam KM,Yip CW,Tse LW,et al.Utility of Mycobacterial interspersed repetitive unit typing for differentiating multidrug-resistant Mycobacterium tuberculosis isolates of the Beijing family[J].J Clin Microbiol,2005,43(1):306-313.DOI:10.1128/jcm.43.1.306-313.2005
[21]Zhang D,An J,Wang YF,et al.Genetic diversity of multidrug-resistant tuberculosis in a resource-limited region of China[J].Int J Infect Dis,2014,29:7-11.DOI:10.1016/j.ijid.2014.05.020
[22]Zhang D,An J,Wang JM,et al.Molecular typing and drug susceptibility of Mycobacterium tuberculosis isolates from Chongqing Municipality,China[J].Infect Genet Evol,2013,13:310-316.DOI:10.1016/j.meegid.2012.10.008
[23]Zhao MH,Liu Y,Liu KM,et al.Genetic Diversity and Drug Resistance of 133 Mycobacterium tuberculosis Isolates from Jiangxi Province,China[J].Mol Biol,2016,5:157.DOI:10.4172/2168-9547.1000157
[24]Dong HY,Liu ZG,Lv B,et al.Spoligotypes of Mycobacterium tuberculosis from different provinces of China[J].J Clin Microbiol,2010,48(11):4102-4106.DOI:0.1128/jcm.00549-10
[25]Pang Y,Zhou Y,Zhao B,et al.Spoligotyping and drug resistance analysis of Mycobacterium tuberculosis strains from national survey in China[J].PLoS ONE,2012,7(3):e32976.DOI:10.1371/journal.pone.0032976
[26]Van SD,Qian L,De Haas PE,et al Predominance of a single genotype of Mycobacterium tuberculosis in countries of East Asia[J].J Clin Microbiol,1995,33(12):3234-3238.
[27]Weniger T,Krawczyk J,Supply P,et al.MIRU-VNTR plus:a web tool for polyphasic genotyping of Mycobacterium tuberculosis complex bacteria[J].Nucleic Acids Res,2010,38:326-331.DOI:10.1093/nar/gkq351
[28]Brudey K,Driscoll JR,Rigouts L,et al.Mycobacterium tuberculosis complex genetic diversity:mining the fourth international spoligotyping database(SpolDB4)for classifcation,population genetics and epidemiology[J].BMC Microbiol,2006,6(23).DOI:10.1186/1471-2180-6-23
[29]Demay C,Liens B.Burguiere T,et al.SITVITWEB—apublicly available international multimarker database for studying Mycobacterium tuberculosis genetic diversity and molecular epidemiology[J].Infect Genet Evol,2012,12(4):755-766.DOI:10.1016/j.meegid.2012.02.004
[30]Bhanu,NV,Soolingen DV,Embden JD,et al.Predominance of a novel Mycobacterium tuberculosis genotype in the Delhi region of India[J].Tuberculosis,2002,82:105-112.DOI:10.1054/tube.2002.0332
[31]Singh UB,Suresh N,Bhanu NV,et al.Predominant tuberculosis spoligotypes,Delhi,India[J].Emerg Infect Dis,2004,10:1138-1142.DOI:10.3201/eid1006.030575
[2]王晓萌,刘志广,柳正卫,等.70株浙江省结核分枝杆菌临床分离株Spoligotyping基因分型研究[J].中国预防医学杂志,2008,9(11):946-949.DOI:10.3969/j.issn.1009-6639.2008.11.003
[3]王晓萌,刘志广,柳正卫,等.Spoligotyping和MLVA用于71株浙江省结核分枝杆菌临床分离株基因分型的初步研究[J].中国人兽共患病学报,2008,24(12):1090-1094.DOI:10.3969/j.issn.1002-2694.2008.12.002
[4]Kamerbeek J,Schouls L,Kolk A,et al.Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology[J].J Clin Microbiol,1997,35:907-914.
[5]Van Soolingen D.Molecular epidemiology of tuberculosis and other mycobacterial infections:main methodologies and achievements[J].J Internal Med,2001,249:1-26.DOI:10.1046/j.1365-2796.2001.00772.x
[6]Sola C,Filliol I,Legrand E,et al.Genotyping of the Mycobacterium tuberculosis complex using MIRUs:association with VNTR and spoligotyping for molecular epidemiology and evolutionary genetics[J].Infect Genet Evol,2003,3:125-133.DOI:10.1016/s1567-1348(03)00011-x
[7]杨修军,万康林,王艳华,等.应用Spoligotyping技术对吉林省结核分枝杆菌基因分型的研究[J].中国卫生工程学,2008,7(6):326-328.DOI:10.3969/j.issn.1671-4199.2008.06.002
[8]刘飞鹰,刘志广,王喜文,等.Spoligotyping对广西地区208株结核分枝杆菌临床分离株的基因分型[J].中国人兽共患病学报,2007,23(12):1226-1230.DOI:10.3969/j.issn.1002-2694.2007.12.015
[9]刘洁,同重湘,李君莲,等.中国西北部两省区结核分枝杆菌临床分离株Spoligotyping分型初步分析[J].实用预防医学,2014,21(5):522-525.DOI:10.3969/j.issn.1006-3110.2014.05.004
[10]Liu J,Tong CX,Liu J,et al.First insight into the genotypic diversity of clinical Mycobacterium tuberculosis isolates from Gansu province,China[J].PLoS ONE,2014,9(6):e99357.DOI:10.1371/journal.pone.0099357
[11]杨筠,董海燕,李定越,等.四川省106株结核分枝杆菌spoligotyping基因分型研究[J].医学研究杂志,2012,41(5):28-31.DOI:10.3969/j.issn.1673-548X.2012.05.010
[12]Jiao WW,Mokrousov I,Sun GZ,et al.Evaluation of new variable-number tandem-repeat systems for typing Mycobacterium tuberculosis with Beijing genotype isolates from Beijing,China[J].J Clin Microbiol,2008,46(3):1045-1049.DOI:10.1128/jcm.01869-07
[13]Wang J,Liu Y,Zhang CL,et al.Genotypes and characteristics of clustering and drug susceptibility of Mycobacterium tuberculosis isolates collected in Heilongjiang province,China[J].J Clin Microbiol,2011,49(4):1354-1362.DOI:10.1128/jcm.02274-10
[14]Liu Q,Yang DD,Xu WG,et al.Molecular typing of mycobacterium tuberculosis isolates circulating in Jiangsu Province,China[J].BMC Infectious Diseases,2011,11:288.DOI:10.1186/1471-2334-11-288
[15]Lu W,Lv B,Liu Q,et al.Genotypes of Mycobacterium tuberculosis isolates in rural China:Using MIRU-VNTR and spoligotyping methods[J].Scand J Infect Dis,2014,46:98-106.DOI:10.3109/00365548.2013.858182
[16]Yu Q,Su YK,Lu B,et al.Genetic diversity of Mycobacterium tuberculosis isolates from inner mongolia,China[J].PLoS ONE,2013,8(5):e57660.DOI:10.1371/journal.pone.0057660
[17]Ma X,Wang HY,Deng YF,et al.rpoB gene mutations and molecular characterization of rifampin-resistant Mycobacterium tuberculosis isolates from Shandong province,China[J].J Clin Microbiol,2006,44(9):3409-3412.DOI:10.1128/jcm.00515-06
[18]Han H,Wang F,Xiao Y,et al.Utility of mycobacterial interspersed repetitive unit typing for differentiating Mycobacterium tuberculosis isolates in Wuhan,China[J].J Med Microbiol,2007,56,1219-1223.DOI:10.1099/jmm.0.47005-0
[19]Dong H,Shi L,Zhao X,et al.Genetic Diversity of Mycobacterium tuberculosis Isolates from Tibetans in Tibet,China[J].PLoS ONE,2012,7(3):e33904.DOI:10.1371/journal.pone.0033904
[20]Kam KM,Yip CW,Tse LW,et al.Utility of Mycobacterial interspersed repetitive unit typing for differentiating multidrug-resistant Mycobacterium tuberculosis isolates of the Beijing family[J].J Clin Microbiol,2005,43(1):306-313.DOI:10.1128/jcm.43.1.306-313.2005
[21]Zhang D,An J,Wang YF,et al.Genetic diversity of multidrug-resistant tuberculosis in a resource-limited region of China[J].Int J Infect Dis,2014,29:7-11.DOI:10.1016/j.ijid.2014.05.020
[22]Zhang D,An J,Wang JM,et al.Molecular typing and drug susceptibility of Mycobacterium tuberculosis isolates from Chongqing Municipality,China[J].Infect Genet Evol,2013,13:310-316.DOI:10.1016/j.meegid.2012.10.008
[23]Zhao MH,Liu Y,Liu KM,et al.Genetic Diversity and Drug Resistance of 133 Mycobacterium tuberculosis Isolates from Jiangxi Province,China[J].Mol Biol,2016,5:157.DOI:10.4172/2168-9547.1000157
[24]Dong HY,Liu ZG,Lv B,et al.Spoligotypes of Mycobacterium tuberculosis from different provinces of China[J].J Clin Microbiol,2010,48(11):4102-4106.DOI:0.1128/jcm.00549-10
[25]Pang Y,Zhou Y,Zhao B,et al.Spoligotyping and drug resistance analysis of Mycobacterium tuberculosis strains from national survey in China[J].PLoS ONE,2012,7(3):e32976.DOI:10.1371/journal.pone.0032976
[26]Van SD,Qian L,De Haas PE,et al Predominance of a single genotype of Mycobacterium tuberculosis in countries of East Asia[J].J Clin Microbiol,1995,33(12):3234-3238.
[27]Weniger T,Krawczyk J,Supply P,et al.MIRU-VNTR plus:a web tool for polyphasic genotyping of Mycobacterium tuberculosis complex bacteria[J].Nucleic Acids Res,2010,38:326-331.DOI:10.1093/nar/gkq351
[28]Brudey K,Driscoll JR,Rigouts L,et al.Mycobacterium tuberculosis complex genetic diversity:mining the fourth international spoligotyping database(SpolDB4)for classifcation,population genetics and epidemiology[J].BMC Microbiol,2006,6(23).DOI:10.1186/1471-2180-6-23
[29]Demay C,Liens B.Burguiere T,et al.SITVITWEB—apublicly available international multimarker database for studying Mycobacterium tuberculosis genetic diversity and molecular epidemiology[J].Infect Genet Evol,2012,12(4):755-766.DOI:10.1016/j.meegid.2012.02.004
[30]Bhanu,NV,Soolingen DV,Embden JD,et al.Predominance of a novel Mycobacterium tuberculosis genotype in the Delhi region of India[J].Tuberculosis,2002,82:105-112.DOI:10.1054/tube.2002.0332
[31]Singh UB,Suresh N,Bhanu NV,et al.Predominant tuberculosis spoligotypes,Delhi,India[J].Emerg Infect Dis,2004,10:1138-1142.DOI:10.3201/eid1006.030575