Detection of Aphanomyces astaci in North American crayfish by polymerase chain reaction

Diseases of Aquatic Organisms

Annabelle Culas¹  Birgit Oidtmann¹  Peter Steinbauer¹  Rudolf W. Hoffmann¹  Sheila Geiger¹ 

关键词: Aphanomyces astaci;    Oomycetes;    Detection;    Polymerase chain reaction;    North American crayfish;    Orconectes limosus;    Pacifastacus leniusculus;    Internal transcribed spacer region;   

DOI  :  10.3354/dao072053

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: We present a PCR based method to detect Aphanomyces astaci in North American crayfish. Primers were designed to specifically amplify parts of the internal transcribed spacer (ITS) regions and the 5.8 rRNA gene of A. astaci. A single round and a semi-nested assay were tested for their sensitivity and specificity. Specificity of the PCR assays was tested against several closely related Aphanomyces species, other Oomycetes and some non-A. astaci DNA that might be found in or on crayfish. The single round assay was fully specific against all DNA tested. In the semi-nested assay, cross-reaction was seen when the equivalent of 40000 or more genomic units of A. invadans or A. frigidophilus were entered into the PCR reaction. The lower detection limit of both assays lies around 1 genomic unit of A. astaci. Investigation of various parts of the exoskeleton of 3 North American crayfish species revealed that for O. limosus and P. leniusculus the telson and soft abdominal cuticle yielded a positive PCR reaction most frequently. For the third species, Procambarus clarkii, only 1 individual tested positive, so no conclusion as to preferred infestation site(s) could be drawn.

【 授权许可】

Unknown