Abstract:To understand the characteristics of genotype in Vibrio parahaemolyticus isolates and its correlation with the drug resistance, we collected 166 samples from different sources in Jiangsu Province from 2006 to 2014. In this study, three virulence genes including thermoliable hemolsin gene (tlh), thermostable direct hemolysin gene (tdh), TDH-related hemolysin gene (trh) and the pandemic-specific markers (orf8) and the outer membrane protein coding gene (vpa0166) were amplified by polymerase chain reaction (PCR), the drug resistance of 8 commonly used antibiotics was detected by the Kirby-Bauer method (KB). According to the sequencing results of PCR products for vpa0166 gene, all isolates were divided into two genotypes, in which, the nucleotide sequence homology between type-A and type-B was 94%, i.e., the size of type-A (996 bp) was 54 bases lower than type-B (1 050 bp). Moreover, partial deletion of vpa0166 gene significantly enhanced the susceptibility to ciprofloxacin, and the P value was <0.05. It is concluded that the diversity of vpa0166 gene is closely correlated with ciprofloxacin resistance.
周海波,薛峰,高璐,饶胜其,杨振泉. 副溶血性弧菌vpa0166基因与耐药性相关性研究[J]. 中国人兽共患病学报, 2018, 34(6): 509-514.
ZHOU Hai-bo, XUE Feng, GAO Lu, RAO Sheng-qi, YANG Zhen-quan. Relationship between vpa0166 gene and drug resistance of Vibrio parahemolyticus. Chinese Journal of Zoonoses, 2018, 34(6): 509-514.
[1] Wu Y, Wen J, Ma Y, et al.Epidemiology of foodborne disease outbreaks caused by Vibrio parahaemolyticus, China, 2003-2008[J].Food Control, 2014, 46(46): 197-202.DOI: 10.1016/j.foodcont.2014.05.023 [2] Bej AK, Patterson DP, Brasher CW, et al.Detection of total and hemolysin-producing Vibrio parahaemolyticus, in shellfish using multiplex PCR amplification of tl, tdh, and trh[J].J Microbiol Methods, 1999, 36(3): 215-225.DOI: 10.1016/S0167-7012(99)00037-8 [3] Shirai H, Ito H, Hirayama T, et al.Molecular epidemiologic evidence for association of thermostable direct hemolysin (TDH) and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis[J].Infect Immun, 1990, 58(11): 3568-3573. [4] Okuda J, Ishibashi M, Hayakawa E, et al.Emergence of a unique O3∶K6 clone of Vibrio parahaemolyticus in Calcutta, India, and isolation of strains from the same clonal group from Southeast Asian travelers arriving in Japan[J].J Clin Microbiol, 1997, 35(12): 3150-3155. [5] Nasu H, Iida T, Sugahara T, et al.A filamentous phage associated with recent pandemic Vibrio parahaemolyticus O3∶K6 strains[J].J Clin Microbiol, 2000, 38(6): 2156-2161. [6] Kitiyodom S, Khemtong S, Wongtavatchai J, et al.Characterization of antibiotic resistance in Vibrio spp.isolated from farmed marine shrimps (Penaeus monodon)[J].FEMS Microbiol Ecol, 2010, 72(2): 219-227.DOI: 10.1111/j.1574-6941.2010.00846.x [7] 戎建荣, 李连青, 王淑峰, 等.外膜孔道蛋白ompC、ompF的表达与大肠埃希菌耐药的相关性研究[J].中华医院感染学杂志, 2009, 19(6): 621-624.DOI:10.3321/j.issn:1005-4529.2009.06.007. [8] Sato T, Yokota S, Okubo T, et al.Contribution of the AcrAB-TolC efflux pump to high-level fluoroquinolone resistance in Escherichia coli isolated from dogs and humans[J].J Vet Med Sci, 2013, 75(4): 407-414.DOI:10.1292/jvms.12-0186 [9] 石亚素, 张行钦, 薛超波, 等.舟山海产品副溶血性弧菌污染及毒力基因分析[J].中国公共卫生, 2007, 23(9): 1135-1136.DOI:10.3321/j.issn:1001-0580.2007.09.058 [10] Tada J, Ohashi T, Nishimura N, et al.Detection of the thermostable direct hemolysin gene (tdh) and thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus by polymerase chain reaction[J].Mol Cell Probes, 1992, 6(6): 477-487.DOI: 10.1016/0890-8508(92)90044-X [11] M100-S20, performance standards for antimicrobial susceptibility testing: nineteenth informational supplement[S].Wayne, PA: Clinical and Laboratory Standards Institute, 2010. [12] Sarkar BL, Nair GB, Sircar BK, et al.Incidence and level of Vibrio parahaemolyticus associated with freshwater plankton[J].Appl Environ Microbiol, 1983, 46(1): 288-290. [13] Theethakaew C, Feil EJ, Castilloramírez S, et al.Genetic relationships of Vibrio parahaemolyticus isolates from clinical, human carrier, and environmental sources in Thailand, determined by multilocus sequence analysis[J].Appl Environ Microbiol, 2013, 79(7): 2358-2370.DOI: 10.1128/AEM.03067-12 [14] Velazquezroman J, Leónsicairos N, Floresvillaseor H, et al.Pandemic Vibrio parahaemolyticus O3∶K6 present in the Coastal Environment of Northwest Mexico is associated with recurrent diarrheal cases from 2004 to 2010[J].Appl Environ Microbiol, 2012, 78(6): 1794-1803.DOI: 10.1128/AEM.06953-11 [15] Okura M, Osawa R, Iguchi A, et al.Genotypic analyses of Vibrio parahaemolyticus and development of a pandemic group-specific multiplex PCR assay[J].J Clin Microbiol, 2003, 41(10): 4676-4782.DOI: 10.1128/JCM.41.10.4676-4682.2003 [16] 李薇薇, 梅玲玲, 唐震, 等.2007-2009年中国副溶血性弧菌临床分离株分子特征分析[J].中华预防医学杂志, 2014, 48(1): 44-52.DOI:10.3760/cma.j.issn.0253-9624.2014.01.010. [17] 黄锐敏, 陈辉, 袁月明.2004-2006年深圳南山区副溶血性弧菌菌群菌型分布及耐药分析[J].中国卫生检验杂志, 2007, 17(7): 1275-1276.DOI:10.3969/j.issn.1004-8685.2007.07.056. [18] 谭海芳, 丁丽娜, 林凤, 等.肇庆市副溶血性弧菌血清分型和耐药性研究[J].实用预防医学, 2010, 17(2): 349-352.DOI:10.3969/j.issn.1006-3110.2010.02.061 [19] 秦思, 沈赟, 马恺, 等.2012年江苏省食源性致病菌耐药监测分析[J].江苏预防医学, 2014, 25(1): 28-30.DOI:10.3969/j.issn.1006-9070.2014.01.010 [20] 吕虹, 李莉, 黄玉兰, 等.2006-2013年四川省副溶血性弧菌毒力基因及耐药分析[J].中国卫生检验杂志, 2015, 25(12): 2045-2048. [21] Duval V, Nicoloff H, Levy SB.Combined inactivation of lon and ycgE decreases multidrug susceptibility by reducing the amount of OmpF porin in Escherichia coli[J].Antimicrob Agents Chemother, 2009, 53(11): 4944-4948.DOI: 10.1128/AAC.00787-09 [22] Schneiders T, Amyes SGB, Levy SB.Role of AcrR and RamA in fluoroquinolone resistance in clinical Klebsiella pneumoniae isolates from Singapore[J].Antimicrob Agents Chemother, 2003, 47(9): 2831-2837.DOI: 10.1128/AAC.47.9.2831-2837.2003 [23] Henrichfreise B, Wiegand I, Pfister W, et al.Resistance mechanisms of multiresistant Pseudomonas aeruginosa strains from Germany and correlation with hypermutation[J].Antimicrob Agents Chemother, 2007, 51(11): 4062-4070.DOI: 10.1128/AAC.00148-07