Chanu Khangembam Victoria, Thakuria Dimpal, Pant Vinita, Bisht Sweta, Tandel Ritesh Shantilal
ICAR-Directorate of Coldwater Fisheries Research, Bhimtal, Uttarakhand-263136, India.
Biotechnol Rep (Amst). 2022 Aug 9;35:e00758. doi: 10.1016/j.btre.2022.e00758. eCollection 2022 Sep.
is the most important pathogen under the genus, which causes devastating oomycete diseases in freshwater fish. At present, the most common molecular method for identification of species is sequencing of ribosomal DNA internal transcribed spacer (rDNA-ITS) region. In this study, a highly sensitive multiplex PCR targeting rDNA-ITS region and a hypothetical protein gene was developed using two sets of primer pair. In this PCR, two amplicons of different size of 750 bp and 365 bp are produced only in case of while other species had single amplicon. This protocol could also differentiate species from other fungus based on the size of rDNA-ITS region. The protocol does not require sequencing and can identify in a single reaction. Therefore, the multiplex PCR developed in this study may prove to be an easier, faster and cheaper molecular method for identification of
是该属中最重要的病原体，可导致淡水鱼毁灭性的卵菌病。目前，鉴定该物种最常用的分子方法是核糖体DNA内转录间隔区（rDNA-ITS）区域测序。在本研究中，使用两组引物对开发了一种针对rDNA-ITS区域和一个假定蛋白基因的高灵敏度多重PCR。在该PCR中，仅在存在该物种时会产生两个大小分别为750 bp和365 bp的不同扩增子，而其他物种只有单一扩增子。该方法还可根据rDNA-ITS区域的大小将该物种与其他真菌区分开来。该方法无需测序，可在单一反应中鉴定该物种。因此，本研究中开发的多重PCR可能是一种更简便、快速且廉价的用于鉴定该物种的分子方法。
