Development and application of reverse transcription loop-mediated isothermal amplification for detection of IHNV 
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1. Heilongjiang Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin 150070, China; 2. Benxi Agrimarine Industries Inc., Benxi of Liaoning Province 117000, China

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s941

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    Abstract:

    A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was developed for detection of infectious haematopoietic necrosis virus (IHNV). A set of specific primers were designed based on the IHNV L-protein sequence. IHNV RNA was used as a template for RT-LAMP in a reaction mixture containing reverse transcriptase and Bst DNA polymerase. The RT-LAMP products were observed by the naked eye by adding SYBR Green I fluorescent dye. Positive samples exhibited a fluorescent green color whereas negative samples exhibited a reddish brown color. The RT-LAMP assay had no cross reactivity with infectious pancreatic necrosis virus (IPNV) and viral haemorrhagic septicaemia virus (VHSV). The RT-LAMP assay was highly sensitive, and could detect < 10 pfu IHNV RNA. We confirmed these results using 2% agarose gel electrophoresis. In summary, we developed a sensitive, specific, low cost, and simple RT-LAMP method for rapid field diagnosis of IHNV. The method can achieve high throughput detection, without relying on any special equipment.

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刘淼,徐黎明,卢彤岩,赵景壮,曹永生,刘红柏,尹家胜,张金凤,冯剑. 鱼类传染性造血器官坏死病毒 RT-LAMP 检测方法的建立及应用 [J]. Jounal of Fishery Sciences of China, 2014,[volume_no](5):1065-1071

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  • Online: August 04,2015
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