Molecular characteristics of clinical isolates of Vibrio parahaemolyticus serogroup O4 in Hangzhou, Zhejiang, 2013−2018
-
摘要:
目的 分析浙江省杭州市腹泻样本中O4血清群副溶血弧菌分子特征。 方法 对2013—2018年杭州市腹泻样本中分离的1 061株副溶血弧菌进行血清凝集分析，采用qPCR方法检测其中147株O4血清群副溶血弧菌的耐热直接溶血素（tdh）、耐热相关溶血素（trh）、不耐热溶血素（tlh）和toxRS/new基因，并用脉冲场凝胶电泳（PFGE）方法进行分子分型。 结果 血清型为O4∶KUT（87.07%, 128/147）、O4∶K8（10.20%, 15/147）和O4∶K9（2.72%, 4/147）的菌株均为tdh+trh-tlh+toxRS/new-基因型。 根据PFGE图谱，菌株可以分成PA（以2016年后分离的菌株为主）和PB（2013—2018年菌株）2个簇，分别包含15和16个不同带型。 其中O4∶K8菌株均处于PA簇，O4∶K9菌株均处于PB簇，O4∶KUT菌株分布于PA和PB簇中。 结论 近年来杭州市分离自腹泻患者的O4∶KUT血清型菌株存在2个流行克隆群。 其中PB簇O4∶KUT菌株持续存在，PA簇O4∶KUT菌株在2016年出现并引起本地散在暴发，该型别菌株有成为地区流行克隆群的可能。 Abstract:Objective To analyze molecular characteristics of Vibrio parahaemolyticus serogroup O4 in diarrhea patients in Hangzhou, Zhejiang province. Methods A total of 1 061 V. parahaemolyticus strains isolated from samples of diarrhea cases in Hangzhou from 2013 to 2018 were serotyped by serum agglutination, and 147 isolates of V. parahaemolyticus serogroup O4 were used to detect the virulence genes of thermostable direct hemolysin (tdh), thermostable hemolysin (trh), thermolabile hemolysin (tlh), toxRS/new genes by qPCR, and pulse-field gel electrophoresis (PFGE) was used. Results The serogroup O4 included O4∶KUT (87.07%, 128/147), O4∶K8 (10.20%, 15/147) and O4∶K9 (2.72%, 4/147). The qPCR results showed that the virulence genes carried by the strains were all tdh+trh-tlh-toxRS/new-. The PFGE patterns had two main clusters, PA (mainly isolates after 2016) and PB (isolates from 2013 to 2018), which contained 15 and 16 patterns respectively. All the O4∶K8 strains belonged to PA cluster, and O4∶K9 strains belonged to PB cluster. Conclusion There were two different O4∶ KUT clones in local food-borne isolates in Hangzhou in recent years. The O4∶KUT strains of pattern PB have been detected since 2013. The O4∶KUT strains of pattern PA, which were detected in 2016, have caused food-borne outbreaks since then and have the potential to become a local epidemic clone. -
图 1 147株副溶血弧菌的脉冲场凝胶电泳图谱聚类结果
Figure 1. Dendrogram of PFGE patterns of 147 V. parahaemolyticus isolates
表 1 2013－2018年杭州地区腹泻患者来源副溶血弧菌血清型分布（株）
Table 1. Serotype distribution of V. parahaemolyticus isolated from diarrhea patients in Hangzhou, 2013−2018
年份 O3∶K6 O4∶KUT O4∶K8 O4∶K9 其他 总计 2013 43 19 10 4 9 85 2014 38 17 1 0 9 65 2015 73 6 1 1 14 95 2016 183 126 18 0 31 358 2017 151 52 12 0 15 230 2018 149 25 15 11 28 228 合计 637 245 57 16 106 1 061 
下载: 导出CSV
表 2 2013－2018年O3∶K6和O4∶KUT菌株来源患者的年龄和性别分布
Table 2. Sex and age distribution of patients infected with V. parahemolyticus O3∶K6 and O4∶KUT serotype, 2013−2018
年龄组（岁） O3∶K6菌株来源病例数 O4∶KUT菌株来源病例数 男性 女性 总计 男性 女性 总计 <7 1 3 4 0 0 0 7～ 16 9 25 7 3 10 19～ 216 213 429 104 66 170 41～ 60 78 138 21 23 44 >60 14 27 45 5 16 21 合计 307 330 637 137 108 245
下载: 导出CSV
-
[1] Joseph SW, Colwell RR, Kaper JB. Vibrio parahaemolyticus and related halophilic Vibrios[J]. CRC Crit Rev Microbiol, 1982, 10(1): 77–124. DOI: 10.3109/10408418209113506. [2] Li YP, Xie TF, Pang R, et al. Food-borne Vibrio parahaemolyticus in China: prevalence, antibiotic susceptibility, and genetic characterization[J]. Front Microbiol, 2020, 11: 1670. DOI: 10.3389/fmicb.2020.01670. [3] Letchumanan V, Chan KG, Lee LH. Vibrio parahaemolyticus: a review on the pathogenesis, prevalence, and advance molecular identification techniques[J]. Front Microbiol, 2014, 5: 705. DOI: 10.3389/fmicb.2014.00705. [4] Centers for Disease Control and Prevention (CDC). Vibrio illnesses after Hurricane Katrina-multiple states, August-September 2005[J]. MMWR Morb Mortal Wkly Rep, 2005, 54(37): 928–931. [5] Centers for Disease Control and Prevention. Vibrio species causing vibriosis[EB/OL]. (2019-03-05)[2020-08-28]. https://www.cdc.gov/vibrio/index.html. [6] Liu JK, Bai L, Li WW, et al. Trends of foodborne diseases in China: lessons from laboratory-based surveillance since 2011[J]. Front Med, 2018, 12(1): 48–57. DOI: 10.1007/s11684−017−0608−6. [7] Paudyal N, Pan H, Liao XY, et al. A meta-analysis of major foodborne pathogens in Chinese food commodities between 2006 and 2016[J]. Foodborne Pathog Dis, 2018, 15(4): 187–197. DOI: 10.1089/fpd.2017.2417. [8] Espejo RT, García K, Plaza N. Insight into the origin and evolution of the Vibrio parahaemolyticus pandemic strain[J]. Front Microbiol, 2017, 8: 1397. DOI: 10.3389/fmicb.2017.01397. [9] Ribot EM, Fair MA, Gautom R, et al. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157∶H7, Salmonella, and Shigella for PulseNet[J]. Foodborne Pathog Dis, 2006, 3(1): 59–67. DOI: 10.1089/fpd.2006.3.59. [10] Wong HC, Liu SH, Chiou CS, et al. A pulsed-field gel electrophoresis typing scheme for Vibrio parahaemolyticus isolates from fifteen countries[J]. Int J Food Microbiol, 2007, 114(3): 280–287. DOI: 10.1016/j.ijfoodmicro.2006.09.024. [11] Hunter SB, Vauterin P, Lambert-Fair MA, et al. Establishment of a universal size standard strain for use with the PulseNet standardized pulsed-field gel electrophoresis protocols: converting the national databases to the new size standard[J]. J Clin Microbiol, 2005, 43(3): 1045–1050. DOI: 10.1128/JCM.43.3.1045−1050.2005. [12] Nair GB, Ramamurthy T, Bhattacharya SK, et al. Global dissemination of Vibrio parahaemolyticus serotype O3: K6 and its serovariants[J]. Clin Microbiol Rev, 2007, 20(1): 39–48. DOI: 10.1128/CMR.00025−06. [13] Matsumoto C, Okuda J, Ishibashi M, et al. Pandemic spread of an O3: K6 clone of Vibrio parahaemolyticus and emergence of related strains evidenced by arbitrarily primed PCR and toxRS sequence analyses[J]. J Clin Microbiol, 2000, 38(2): 578–585. DOI: 10.1128/JCM.38.2.578−585.2000. [14] 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): 4676–4682. DOI: 10.1128/JCM.41.10.4676−4682.2003. [15] Wang LP, Zhou SX, Wang X, et al. Etiological, epidemiological, and clinical features of acute diarrhea in China[J]. Nat Commun, 2021, 12(1): 2464. DOI: 10.1038/s41467−021−22551−z. [16] Su YC, Liu CC. Vibrio parahaemolyticus: a concern of seafood safety[J]. Food Microbiol, 2007, 24(6): 549–558. DOI: 10.1016/j.fm.2007.01.005. [17] Han DS, Tang H, Ren CL, et al. Prevalence and genetic diversity of clinical Vibrio parahaemolyticus isolates from China, revealed by multilocus sequence typing scheme[J]. Front Microbiol, 2015, 6: 291. DOI: 10.3389/fmicb.2015.00291. [18] Li P, Xin WW, Kang L, et al. Genetic and population analyses of Vibrio parahaemolyticus isolates from three major coastal regions in China[J]. Future Microbiol, 2018, 13(11): 1261–1269. DOI: 10.2217/fmb−2018−0060. [19] Yang C, Pei XY, Wu YR, et al. Recent mixing of Vibrio parahaemolyticus populations[J]. ISME J, 2019, 13(10): 2578–2588. DOI: 10.1038/s41396−019−0461−5. [20] Li RP, Wang J, Wang L, et al. Genetic and serological characterization of capsular antigen untypeable Vibrio parahaemolyticus strains reveal novel K serotypes and epidemiological characteristics in Shandong, China[J]. Int J Food Microbiol, 2021, 347: 109188. DOI: 10.1016/j.ijfoodmicro.2021.109188. [21] 张巍巍, 李颖, 王群, 等. 全基因组数据应用于多克隆副溶血弧菌感染暴发的病原学分析[J]. 疾病监测,2020,35(6):518–522. DOI:10.3784/j.issn.1003−9961.2020.06.013.Zhang WW, Li Y, Wang Q, et al. Application of whole genome sequencing in etiology investigation of an outbreak of polyclonal Vibrio parahaemolyticus infection[J]. Dis Surveill, 2020, 35(6): 518–522. DOI: 10.3784/j.issn.1003−9961.2020.06.013. [22] 杨元斌, 高红, 章丹阳, 等. 一起食源性疾病暴发中的副溶血性弧菌病原学分析[J]. 疾病监测,2017,32(6):481–485. DOI:10.3784/j.issn.1003−9961.2017.06.010.Yang YB, Gao H, Zhang DY, et al. Etiology of a Vibrio parahemolyticus caused foodborne-disease outbreak[J]. Dis Surveill, 2017, 32(6): 481–485. DOI: 10.3784/j.issn.1003−9961.2017.06.010. [23] Wang HL, Tang XY, Su YC, et al. Characterization of clinical Vibrio parahaemolyticus strains in Zhoushan, China, from 2013 to 2014[J]. PLoS One, 2017, 12(7): e0180335. DOI: 10.1371/journal.pone.0180335. -
点击查看大图
图(1) / 表(2)
计量
- 文章访问数: 88
- HTML全文浏览量: 51
- PDF下载量: 19
- 被引次数: 0