Re-examination of the prevalence of Aphanomyces astaci in North American crayfish populations in Central Europe by TaqMan MGB real-time PCR

Abstract

We applied quantitative TaqMan minor groove binder real-time polymerase chain reaction (PCR) on DNA isolates from soft abdominal cuticle of 460 North American crayfish Orconectes limosus and Pacifastacus leniusculus, previously tested for Aphanomyces astaci presence by conventional semi-nested PCR. The present study shows that previous results based on the semi-nested PCR were not substantially influenced by false positives but might have suffered from some false negatives at low agent levels.

Combining alternative methods may therefore provide more reliable conclusions on the patogen's presence.

Keywords

• Crayfish plague
• Aphanomyces astaci
• Pacifastacus leniusculus
• Orconectes limosus
• Semi-nested PCR
• Real-time PCR
• Pathogen detection
• Agent level