食品科学 ›› 2018, Vol. 39 ›› Issue (23): 16-21.doi: 10.7506/spkx1002-6630-201823003

• 基础研究 • 上一篇    下一篇

人工污染GII.4型诺如病毒在太平洋牡蛎中的蓄积与分布特性

苏来金1,2,3,马丽萍1,刘 慧1,周德庆1,*   

  1. 1.青岛海洋科学与技术试点国家实验室,海洋药物与生物制品功能实验室,中国水产科学研究院黄海水产研究所,山东 青岛 266071;2.上海海洋大学食品学院,上海 201306;3.温州市农业科学研究院,浙江 温州 325006
  • 出版日期:2018-12-15 发布日期:2018-12-17
  • 基金资助:
    国家自然科学基金面上项目(31471663)

Accumulation and Distribution of GII.4 Norovirus in Artificially Contaminated Pacific Oyster

SU Laijin1,2,3, MA Liping1, LIU Hui1, ZHOU Deqing1,*   

  1. 1. Pilot National Laboratory for Marine Science and Technology (Qingdao), Laboratory for Marine Drugs and Biological Products, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; 2. College of Food Sciences and Technology, Shanghai Ocean University, Shanghai 201306, China; 3. Wenzhou Academy of Agricultural Science, Wenzhou 325006, China
  • Online:2018-12-15 Published:2018-12-17

摘要: 牡蛎是食源性诺如病毒传播的重要载体,诺如病毒在牡蛎中的检出呈现明显的季节性和组织分布差异。为了解诺如病毒在牡蛎中的蓄积规律和组织分布,采用实验室人工污染GII.4型诺如病毒并控制环境进行牡蛎养殖,采用实时荧光定量反转录聚合酶链式反应(reverse transcription polymerase chain reaction,RT-PCR)和免疫组织化学方法,检测牡蛎不同组织中诺如病毒的含量和分布。实时荧光定量RT-PCR结果表明,GII.4型诺如病毒在牡蛎外套膜中的含量比较稳定,在(8.75±0.36)×104~(9.12±0.49)×104 拷贝数之间;在牡蛎鳃中的含量先增加后减少,6 h时达到最高((1.15±0.15)×106 拷贝数);在牡蛎消化道中含量逐渐增加,12 h时达(1.06±0.14)×106 拷贝数并趋于稳定;24 h后GII.4型诺如病毒主要分布于牡蛎的消化道组织中;净化对牡蛎体内富集的GII.4型诺如病毒含量无显著影响(P>0.05)。免疫组织化学结果显示,GII.4型诺如病毒广泛分布在牡蛎外套膜边缘、鳃壁和消化道内壁上,其分布随时间变化趋势与实时荧光定量RT-PCR测定结果一致。

关键词: 诺如病毒, 人工污染, 牡蛎, 蓄积, 分布

Abstract: Oysters are one of the most common vectors for the transmission of noroviruses (NoVs), which are one of the most important food-borne viral pathogens worldwide. The distribution of NoVs in oysters varies significantly with seasons and tissues. In order to understand the accumulation and distribution of norovirus in oysters, oysters were farmed under controlled conditions after being artificially contaminated with GII.4 NoV in the laboratory. The contents and distribution of GII.4 NoV in various oyster tissues were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The real-time quantitative RT-PCR results showed that the content of GII.4 NoV remained stable in the range of (8.75 ± 0.36) × 104 to (9.12 ± 0.49) × 104 copies for oyster mantle, but initially increased to a maximum of (1.15 ± 0.15) × 106 copies at 6 h and then decreased for oyster gills. The content of GII.4 NoV in oyster digestive tissues was continuously increased to reach a plateau ((1.06 ± 0.14)×106 copies) at 12 h. GII.4 NoV was mainly accumulated in oyster digestive tissues at 24 h. The content of GII.4 NoV was not significantly changed after purification (P > 0.05). The immunohistochemistry results showed that GII.4 NoV was widely distributed in the mantle edge, the gill wall and the inner wall of digestive tissues. The seasonal distribution of GII.4 NoV in various oyster tissues was in accordance with the real-time quantitative RT-PCR results.

Key words: norovirus, artificial contamination, oyster, accumulation, distribution

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