Page 151 - 《渔业研究》2025年第5期
P. 151

692                                  渔  业  研  究                                     第 47 卷

              可以通过使用一对不等量的引物来高效产生大量                                 franciscana)[J]. Aquaculture, 2003, 220(1−4): 43 − 57.
              ssDNA。而逆转录技术以         RNA  为模板获取      ssDNA,    [  6  ]   赵然,史文军,王李宝,等. 脊尾白虾“僵尸病”的初
              该过程需体外转录,因此该操作受多因素影响。本                                探  [J]. 水产学报,2023,47(9) :099414.
              研究构建的      aPCR  技术与之相比,操作更为简单且                       Zhao R, Shi W J, Wang L B, et al. A preliminary study
              不易污染,能够在         2 h  内获得大量的    ssDNA,且后              on  the  “zombie  disease”  of  Exopalaemon  carinicauda
              续利用该技术可高效、稳定地对实际样品的检测。                                [J]. Journal of Fisheries of China, 2023, 47(9): 099414.
                  本文对比了      qPCR  技术和    HRCA  技术在对实         [  7  ]   Cao  G  N,  Bao  J,  Feng  C  C,  et  al.  First  report  of
              际样本的检测效率。以           qPCR  技术作为检出率的检                  Metschnikowia  bicuspidata  infection  in  Chinese  grass
              测标准,本研究建立的检测中华绒螯蟹二尖梅奇酵                                shrimp  (Palaemonetes  sinensis)  in  China[J].  Trans-
              母的   HRCA  的检出率为      56.25 %,与  qPCR  技术的            boundary  and  Emerging  Diseases,  2022,  69(5):  3133  −
              检出率一致。两种检测方法对实际样本的检测效率                                3141.
              均达到池边检测的要求。                                     [  8  ]   许文军,徐汉祥,金海卫,等. 梭子蟹“乳化病”病原
                                                                    的研究  [J]. 浙江海洋学院学报(自然科学版) ,2003,
               4 结论
                                                                    22(3) :209 − 213.
                  本文建立了检测中华绒螯蟹“牛奶病”病原二                              Xu W J, Xu H X, Jin H W, et al. Study on pathogen of
              尖梅奇酵母的       HRCA  技术。该方法对实际样本检                       “emulsification” disease of Portunus trituberculatus[J].
              测灵敏度为      4.13×10  pg/μL,检出率与     qPCR  技术           Journal  of  Zhejiang  Ocean  University  (Natural  Science
                                1
              一致,具有较高的特异性、重复性和稳定性,且不                                Edition), 2003, 22(3): 209 − 213.
              依赖变温设备,能够满足池边检测的需求,实现较                          [  9  ]   Codreanu  R,  Codreanu-Balcescu  D.  On  two  Metsch-
              为快速检测的目的,可被广泛应用于养殖现场检                                 nikowia yeast species producing hemocoelic infections in
              测,在水产病原检测和病原诊断领域具有很大的应                                Daphnia magna and Artemia salina (Crustacea, Phyllo-
              用潜力。                                                  poda) from Romania[J]. Journal of Invertebrate Patho-
                                                                    logy, 1981, 37(1): 22 − 27.
                  致谢:感谢盘锦光合蟹业有限公司孙娜主
                                                              [10]   Lu C C, Tang K F J, Chen S N. Identification and genet-
              任对中华绒螯蟹的采集提供的指导与帮助。
                                                                    ic  characterization  of  yeasts  isolated  from  freshwater
                                                                    prawns, Macrobrachium rosenbergii de Man, in Taiwan
              参考文献(References) :
                                                                    [J]. Journal of Fish Diseases, 1998, 21(3): 185 − 192.
              [  1  ]   Ding  Z  F.  Current  disease  threats  for  cultivated  crab
                                                              [11]   Zhang  H  Q,  Chi  Z,  Liu  G  L,  et  al.  Metschnikowia  bi-
                   Eriocheir  sinensis  in  China[J].  Transboundary  and
                                                                    cuspidate  associated  with  a  milky  disease  in  Eriocheir
                   Emerging Diseases, 2023, 2023: 3305963.
              [  2  ]   Sun N, Bao J, Liang F, et al. Prevalence of ‘milky dis-  sinensis and its effectitve treatment by Massoia lactone
                                                                    [J]. Microbiological Research, 2021, 242: 126641.
                   ease’ caused by Metschnikowia bicuspidata in Eriocheir
                                                              [12]   Bao J, Chen Y, Xing Y N, et al. Development of a nes-
                   sinensis in Panjin City, China[J]. Aquaculture Research,
                                                                    ted  PCR  assay  for  specific  detection  of  Metschnikowia
                   2022, 53(3): 1136 − 1140.
              [  3  ]   Jiang H B, Bao J, Xing Y N, et al. Proteomic analysis of  bicuspidata infecting Eriocheir sinensis[J]. Frontiers in
                   the  hemolymph  after  Metschnikowia  bicuspidata  infec-  Cellular and Infection Microbiology, 2022, 12: 930585.
                                                              [13]   Xing Y N, Chen Y, Feng C C, et al. Establishment and
                   tion  in  the  Chinese  mitten  crab  Eriocheir  sinensis[J].
                   Frontiers in Immunology, 2021, 12: 659723.       application  of  real-time  fluorescence  quantitative  PCR
              [  4  ]   Bao J, Jiang H B, Shen H B, et al. First description of  detection technology for Metschnikowia bicuspidata dis-
                   milky disease in the Chinese mitten crab Eriocheir sinen-  ease  in  Eriocheir  sinensis[J].  Journal  of  Fungi,  2023,
                   sis  caused  by  the  yeast  Metschnikowia  bicuspidata[J].  9(8): 791.
                   Aquaculture, 2021, 532: 735984.            [14]   Lizardi P M, Huang X H, Zhu Z R, et al. Mutation detec-
              [  5  ]   Moore M M, Strom M S. Infection and mortality by the  tion  and  single-molecule  counting  using  isothermal
                   yeast  Metschnikowia  bicuspidata  var.  bicuspidata  in  rolling-circle  amplification[J].  Nature  Genetics,  1998,
                   Chinook  salmon  fed  live  adult  brine  shrimp  (Artemia  19(3): 225 − 232.
   146   147   148   149   150   151   152   153   154