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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
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