一种用于检测虹鳟（Oncorhynchus mykiss）中传染性造血坏死病毒（IHNV）的环介导等温扩增（LAMP）方法。
A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss).
作者信息
Gunimaladevi I, Kono T, Lapatra S E, Sakai M
机构信息
United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan.
出版信息
Arch Virol. 2005 May;150(5):899-909. doi: 10.1007/s00705-004-0468-7. Epub 2005 Jan 19.
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout (Oncorhynchus mykiss). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 degrees C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a10-fold higher sensitivity was seen for the LAMP protocols.
开发了一种逆转录环介导等温扩增（RT-LAMP）方法，用于检测虹鳟（Oncorhynchus mykiss）中的传染性造血坏死病毒（IHNV）RNA。基于IHNV的G蛋白序列设计了一组四条引物，即用于RT-LAMP和LAMP检测的两条外部引物和两条内部引物。对RT-LAMP和LAMP方法的时间和温度条件进行了优化，均为63℃ 60分钟。发现RT-LAMP和LAMP的检测限相似。当将RT-LAMP和LAMP的灵敏度与传统巢式PCR进行比较时，LAMP方法的灵敏度高10倍。
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