Development of a duplex real-time PCR assay for detection of Perkinsus and Marteilia refringens in shellfish.

畜牧与饲料科学 Pub Date : 2010-09-17 DOI:10.3724/SP.J.1035.2010.00898

Xie Zhixun, Xie Liji, Pang Yaoshan, Liu Jiabo, Deng Xianwen, Xie Zhiqin

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引用次数: 1

Abstract

[Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then shorten the detective cycle.[Method]According to the gene sequence of Perkinsus sp and Marteilia refringens from gene bank, design two pairs of specific primers and two TaqMan probes with different fluorophores labeled. Optimizing the reactive conditions and reagent concentration in order that establishing the duplex real-time PCR method for detecting Perkinsus sp and Marteilia refringens simultaneously.[Result] The sensitivity of the duplex real-time PCR method which about Perkinsus sp and Marteilia refringens is 40 template copies. After combine the templates of Perkinsus sp and Marteilia refringens with different concentrations, this method still could be detect this two protozoan efficiently and synchronously.[Conclusion] The established duplex real-time PCR method for detecting Perkinsus sp. and Marteilia refringens possesses lots of advantages, such as specific, sensitive, rapid, quantitative and reproducible, can be used for clinical detection of infection which was caused by Perkinsus sp. and Marteilia refringens.

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贝类中珀金氏菌和马氏菌双链实时PCR检测方法的建立。

[目的]提高白蜡草和麻豆草的检测准确性和效率，缩短检测周期。[方法]根据基因库中珀金苏斯(Perkinsus sp)和马提利亚(Marteilia refingens)的基因序列，设计两对特异性引物和两种标记不同荧光团的TaqMan探针。优化反应条件和试剂浓度，建立双联实时PCR同时检测珀金苏氏菌和马氏菌的方法。[结果]双链实时荧光定量PCR法对珀金苏和马提兰的检测灵敏度为40个模板拷贝。将不同浓度的珀金苏斯(Perkinsus sp)和瑞金马提利亚(Marteilia refringens)模板组合后，该方法仍能高效、同步地检测出这两种原生动物。[结论]所建立的珀金苏氏菌和麻豆状芽孢杆菌双链实时PCR检测方法具有特异性、敏感性、快速、定量和重复性好等优点，可用于珀金苏氏菌和麻豆状芽孢杆菌感染的临床检测。

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畜牧与饲料科学畜牧业生产与科学研究

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期刊介绍： Animal Husbandry and Feed Science is an academic journal supervised and sponsored by the Academy of Agricultural and Animal Husbandry Sciences of Inner Mongolia Autonomous Region. The journal was founded in 1973, the original name of the journal is “Inner Mongolia Animal Husbandry Science”, changed to the current name in 2003. Domestic uniform issue number: CN 15-1228/S; international standard issue number: ISSN 1672-5190; international journal code (CODEN): XYSKAX. Animal Husbandry and Feed Science is one of the first academic journals recognized by the State Administration of Press, Publication, Radio, Film and Television (SARFT), and has been selected as one of China's scientific and technological core journals (2021 and 2022 editions), RCCSE China's Core Academic Journals (2017-2020), and China's Agricultural and Forestry Core Journals (2020 edition); It is included in the full text of domestic academic resource databases, such as Zhi.com, Wanfang, Wipu, Superstar, etc. The English abstracts of the articles published in the journal are included in the CAB Abstracts database under the Research Center for Agricultural and Biological Sciences of the United Kingdom (CABI) , and in the full text of the database of the U.S. EBSCO; and it is a source journal of the “Annual Report of Impact Factor of Chinese Academic Journals”.

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Development of a duplex real-time PCR assay for detection of Perkinsus and Marteilia refringens in shellfish.