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用于检测与昏睡蟹病(LCD)相关的黑色酵母真菌的特异性引物。

Specific primers for the detection of the black-yeast fungus associated with lethargic crab disease (LCD).

作者信息

Pie Marcio R, Boeger Walter A, Patella Luciana, Vicente Vânia A, Ribeiro Raphael O, Ostrensky Antonio

机构信息

Departamento de Zoologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.

出版信息

Dis Aquat Organ. 2011 Mar 16;94(1):73-5. doi: 10.3354/dao02312.

Abstract

Lethargic crab disease (LCD) is an emerging infirmity that has been causing extensive mortalities in populations of the mangrove land crab Ucides cordatus (Ocypodidae) along the Atlantic coast of Brazil. Previous studies have indicated that LCD is associated with a dematiaceous fungus, Exophiala cancerae de Hoog et al. In the present study, we sequenced the internal transcribed spacer (ITS) of the rDNA region of this black yeast species and developed species-specific PCR primers. Sensitivity tests indicated that the developed protocol is capable of detecting very small amounts of target DNA. Also, the application of the protocol to a variety of other dematiaceous fungi did not generate any false positives. The specific primers provided in the present study represent an important tool for rapidly surveying a large number of crab individuals, as well as environmental samples. Such knowledge will be instrumental in understanding the epidemiological dynamics of LCD.

摘要

嗜睡蟹病(LCD)是一种新出现的疾病,已在巴西大西洋沿岸的红树林招潮蟹(Ucides cordatus,沙蟹科)种群中造成大量死亡。先前的研究表明,LCD与一种暗色真菌——蟹外瓶霉(Exophiala cancerae)有关。在本研究中,我们对这种黑色酵母菌的核糖体DNA(rDNA)区域的内转录间隔区(ITS)进行了测序,并开发了物种特异性PCR引物。敏感性测试表明,所开发的方法能够检测到极少量的目标DNA。此外,将该方法应用于多种其他暗色真菌时未产生任何假阳性结果。本研究中提供的特异性引物是快速检测大量蟹个体以及环境样本的重要工具。这些知识将有助于了解LCD的流行病学动态。

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