吴漆波, 石亚素, 虞艳, 潘军航. 浙江省舟山市售贝类中创伤弧菌的定量检测及毒力基因分析[J]. 疾病监测, 2012, 27(11): 894-897. DOI: 10.3784/j.issn.1003-9961.2012.11.016
引用本文: 吴漆波, 石亚素, 虞艳, 潘军航. 浙江省舟山市售贝类中创伤弧菌的定量检测及毒力基因分析[J]. 疾病监测, 2012, 27(11): 894-897. DOI: 10.3784/j.issn.1003-9961.2012.11.016
WU Qi-bo, SHI Ya-su, YU Yan, PANG Jun-hang. Quantitative analysis and gene typing of Vibrio vulnificus in sea shellfish from Zhoushan market[J]. Disease Surveillance, 2012, 27(11): 894-897. DOI: 10.3784/j.issn.1003-9961.2012.11.016
Citation: WU Qi-bo, SHI Ya-su, YU Yan, PANG Jun-hang. Quantitative analysis and gene typing of Vibrio vulnificus in sea shellfish from Zhoushan market[J]. Disease Surveillance, 2012, 27(11): 894-897. DOI: 10.3784/j.issn.1003-9961.2012.11.016

浙江省舟山市售贝类中创伤弧菌的定量检测及毒力基因分析

Quantitative analysis and gene typing of Vibrio vulnificus in sea shellfish from Zhoushan market

  • 摘要: 目的 了解2010-2012年浙江省舟山市售贝类中创伤弧菌(Vibrio vulnificus,VV)的污染状况与季节性变化规律,及其毒力相关基因分型。 方法 采用3%氯化钠碱性蛋白胨水增菌,纤维二糖一多粘茵素E(cc)琼脂分离贝类海产品中的创伤弧菌,可疑菌落的鉴定采用API20E,采用最近似值法(most probable number,MPN)进行创伤弧菌的定量检测,应用PCR技术进行生物Ⅰ型、Ⅱ型和血清E型鉴定,及vcg、16S rRNA毒力基因分型。 结果 179份样品检出64株创伤弧菌,检出阳性率为35.75%。不同季节检出率差异有统计学意义(2=35.23,P0.05),夏季检出率最高为68%。39株VV菌株为溶血素A基因(hemolysin gene A, vvhA)核酸阳性,1株BT2型,1株血清E型,其余37株均为BT1型。vcgC/E和16S rRNA A/B毒力基因分型结果显示:CB型 31株,CAB型4株,EA型 3株,EAB型 1株。 结论 舟山市售贝类中创伤弧菌的污染状况严重,尤其是夏秋季,毒力基因型呈多样性,以CB型为主。应对贝类的食品安全进行风险评估,预防食源性疾病的暴发。

     

    Abstract: Objective To understand the contamination of Vibrio vulnificus in sea shellfish and the types of related virulence gene in Zhoushan. Methods By using 3% sodium chloride alkaline peptone water, the enrichment of bacteria was conducted, V. vulnificus in sea shellfish was isolated with mCPC Agar Base and suspected colonies were identified by using API20E, the quantitative detection of V. vulnificus was done with most probable number method and PCR was conducted for identification of biotype Ⅰ, type Ⅱ and serotype E of the strains and genotyping of virulence genes of vcg,16srRNA. Results Totally 64 strains of V. vulnificus in 179 samples were detected, the positive rate was 35.75%. The detection rate in summer was highest(68%) and the differences in detection rates in different seasons had statistical significance.(2=35.23,P0.05). Thirty nine strains were hemolysin gene A(vvhA) nucleic acid positive, in which 1 belonged to BT2 type,1 belonged to serotype E and other 37 strains belonged to BT1. The genotyping of virulence gene vcgC/E and 16S rRNA A/B indicated CB type was detected 31 strains, CAB type in 4 strains, EA type in 3 strains and EAB type in 1 strain. Conclusion The contamination of V. vulnificus in sea shellfish was serious in Zhoushan, especially during summer and autumn. The types of virulence genes of the pathogen showed diversity, CB type was predominated. It is necessary to conduct the risk evaluation of sea shellfish food to prevent the outbreak of food borne diseases.

     

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