Loop-mediated isothermal amplification for diagnosing marine pathogens in tissues of Crassostrea spp. and white shrimp, Litopenaeus vannamei, farmed in Mexico

IF 0.5 4区生物学 Q4 MARINE & FRESHWATER BIOLOGY Ciencias Marinas Pub Date : 2021-12-17 DOI:10.7773/cm.v47i4.3095

Ismael Mendoza-Avilés, Carla A Muñoz-Rojas, Mario Rojas, Norma Estrada

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引用次数: 1

Abstract

Loop-mediated isothermal amplification (LAMP) is an accurate, sensitive, rapid, and easy-to-perform method for gene amplification under isothermal conditions, and it has served as a powerful diagnostic tool. In this study, we used LAMP to develop a diagnostic protocol for detecting Vibrio parahaemolyticus and white spot syndrome virus in whiteleg shrimp, and Perkinsus spp. in Crassostrea spp. in Mexico. These pathogens are associated with different diseases and are considered a threat in the aquaculture industry. Infected and uninfected oysters and shrimp were obtained from farms in the northwest coast of Mexico to standardize the LAMP assay. We determined the candidate target genes in the first-round analysis of many sets of primers, and then we chose a set of primers that successfully amplified with Mexican samples. We optimized the LAMP reactions for each pathogen with the chosen primer sets using temperature gradients from 61 to 65 ºC, DNA concentrations from 2.5 pg to 250.0 ng, and reaction times from 10 to 60 min. This study established a diagnostic procedure for detecting pathogens in oysters and shrimp from Mexico. Early diagnosis and treatment of pathogens can immensely reduce disease transmission in aquaculture farms.

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环介导等温扩增用于诊断墨西哥养殖的凡纳滨对虾(Litopenaeus vannamei)和长牡蛎(crasstrea spp.)组织中的海洋病原体

环介导等温扩增（LAMP）是一种准确、灵敏、快速、易于在等温条件下进行基因扩增的方法，是一种强大的诊断工具。在这项研究中，我们使用LAMP开发了一种诊断方案，用于检测墨西哥白腿虾和Crassostrea spp.中的Perkinsus spp.的副溶血性弧菌和白斑综合征病毒。这些病原体与不同的疾病有关，被认为是水产养殖业的威胁。从墨西哥西北海岸的农场获得感染和未感染的牡蛎和虾，以标准化LAMP测定。我们在对多组引物的第一轮分析中确定了候选靶基因，然后我们选择了一组用墨西哥样本成功扩增的引物。我们使用选定的引物组优化了每种病原体的LAMP反应，使用温度梯度为61至65ºC，DNA浓度为2.5 pg至250.0 ng，反应时间为10至60分钟。本研究建立了检测墨西哥牡蛎和虾中病原体的诊断程序。病原体的早期诊断和治疗可以极大地减少水产养殖场的疾病传播。

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来源期刊

Ciencias Marinas 生物-海洋与淡水生物学

CiteScore

1.10

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： A bilingual open-access publication, Ciencias Marinas (CM) is an international peer-reviewed journal that contains original research findings in all areas of marine science. It is published quarterly by the Autonomous University of Baja California, Mexico, and all its contents are publicly available on our journal website. Though a limited number of copies are still printed, the journal is mainly distributed in its electronic format. CM was conceived in 1973 as part of an academic project aimed to entice local researchers to publicly disclose their findings by adopting the culture of peer-review publishing. This academic project evolved into an international journal after accepting papers from researchers in the United States and, eventually, other parts of the world. Because of the diversity in authorship, CM issues were initially published in either Spanish or English, and occasionally in both languages. It was not until 1984 when CM included both language versions of all its contents, and it then became the fully bilingual journal it still is today. At CM we believe our inclusive format allows us not only to address a wider range of submissions from international authors but also to make published findings available to a wider international audience. So whether you are looking for information on the redfish in Icelandic waters or the physical and biological properties of the Gulf of California, feel free to peruse CM contents. You may find them to provide source material for your research.