Development of a multiplex PCR to detect Kudoa spp. and to distinguish Kudoa septempunctata in olive flounder Paralichthys olivaceus
Development of a multiplex PCR to detect Kudoa spp. and to distinguish Kudoa septempunctata in olive flounder Paralichthys olivaceus
复制标题
开发多重 PCR 检测 Kudoa spp。
DOI：
10.1016/j.aquaculture.2016.06.005
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发表时间：
2016
期刊：
影响因子：
4.5
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Shin;S. P. Ishitani;H. Shirakashi;S.
自从被鉴定为人类食物中毒的病原体以来，牙周病已成为橄榄鱼水产养殖业的一个严重问题。为了确保养殖比目鱼的安全，人们建立了多种方法来检测七星鱼。然而，对其他种的检查和检测却没有引起足够的重视。到目前为止，已经有六个Kudoaspp了。据报道，有3种来自树干肌肉，2种来自大脑，1种来自心脏。在对养殖比目鱼的常规检查中，除了先前报道的物种名单外，我们还分离到了几个种，包括库多阿伊格米亚和库多阿·奥瓦伊。本研究建立了检测Kudoaspp基因的多重聚合酶链式反应(MPCR)方法。已知来自日本的橄榄比目鱼。MPCR还能将七星克雷伯菌与其他苦瓜区分开来。同时。建立的mPCR方法成功地检测到7株Kudoaspp。和分化的K。粘孢子的敏感度为103。因此，mPCR提供了一种可靠和相对简单的方法来检测橄榄牙鲆的污染，特别是针对识别七星鱼感染。自从最近发现七星粘虫是引起人类食物中毒的病原体以来，细小的粘虫寄生虫库多阿属(粘虫纲：多瓣纲)的感染已经成为比目鱼养殖中严重关注的问题。尽管已经开发了各种检测方法来检查KK。在七星草属中，对其他种的关注相对较少。我们添加了K。奥瓦瓦和K。Igamito工党的名单。并建立了同时检测Kudoaspp的多重PCR方法。并区分K。七星草属。这为检验Kudoaspp提供了相对简单的方法。在养殖的比目鱼中。
Since the identification ofKudoaseptempunctataas a causative agent of human food poisoning,Kudoainfections have become a serious concern in the aquaculture of olive flounder (Paralichthys olivaceus). In order to ensure the safety of cultured flounder, various methods have been developed to detectK.septempunctata. However, less attention has been paid for inspection and detection of otherKudoaspecies. To date, sixKudoaspp. have been reported from olive flounder, 3 species each from the trunk muscle, 2 species from the brain, and one species from the heart. During our routine inspection of cultured olive flounder, we isolated severalKudoaspecies includingKudoa igamiandKudoa ogawaiin addition to the list of previously reported species. In the present study, a multiplex PCR (mPCR) was developed to detect allKudoaspp. known to date from olive flounder in Japan. The mPCR also differentiateK.septempunctataamong otherKudoaspp. simultaneously. The developed mPCR successfully detected sevenKudoaspp. and differentiatedK. septempunctatawith a sensitivity of 103myxospores. Thus, mPCR offers a reliable and relatively simple method to detectKudoainfection in olive flounder, specifically aiming to identifyK.septempunctatainfection.Statement of RelevanceOlive flounderParalichthys olivaceusis one of the most important fish species for commercial aquaculture and fishery trading in Japan and Korea. Infections of microscopic myxozoan parasite of genusKudoa(Myxozoa: Multivalvulida) has become a serious concern in flounder culture since the recent identification ofKudoa septempunctataas a causative agent of human food poisoning. Although various detection methods have been developed for the inspection ofK. septempunctata, relatively little attentions are given to otherKudoaspp. We addedK. ogawaiandK. igamito the list ofKudoaspp. infecting olive flounder and developed multiplex PCR to simultaneously detectKudoaspp. and distinguishK. septempunctata. This offers relatively simple and method to inspectKudoaspp. in cultured flounder.