A one-step molecular biology method for simple and rapid detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 strain

文献类型： 外文期刊

第一作者： Zeng, W. W.

作者： Zeng, W. W.;Wang, Q.;Wang, Y. Y.;Wu, S. Q.;Xu, D. H.

作者机构：

关键词： diagnosis technique;GCRV;RT-LAMP;segment 6 gene

期刊名称：JOURNAL OF FISH BIOLOGY （ 影响因子：2.051； 五年影响因子：2.25 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： Six reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primers designed against conserved regions of segment 6 (s6) gene were used for the detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 subtype. The entire amplification could be completed within 40 min at 62 center dot 3 degrees C. The RT-LAMP showed higher sensitivity than reverse-transcription polymerase chain reaction (RT-PCR). The RNA detection limit was 10 copies mu l1 for RT-LAMP assay and 100 copies mu l1 for conventional RT-PCR. In specificity tests, no cross-reactivity was detected in other viruses from common aquatic animals. In addition, the reaction results can be visualized by using calcein fluorescent dye. Furthermore, a total of 86 samples were tested by RT-LAMP, RT-PCR and virus isolation. The results demonstrated that all 54 specimens identified as positive by virus isolation were also positive when detected by RT-LAMP. Seven out of 54 samples, however, were misidentified by RT-PCR. The RT-LAMP method is more accurate than conventional RT-PCR. The results indicate that RT-LAMP has potential as a simple and rapid diagnosis technique for the detection of GCRV HZ08 subtype infection.

分类号： Q959.4

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