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

摘要: 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. Both approaches target the internal transcribed spacers pathogen nuclear ribosomal DNA, but apply different specific sequence motifs technologies. The PCR approach seems to provide higher sensitivity; number that positive increased 23 32%, 10 additional populations were indicated as hosting disease agent. However, vast majority newly recorded positives contained very low agent levels, 5 50 PCR-forming units. An isolate producing a false result (apparently undescribed related A. astaci) remained negative using present study shows previous results based not substantially influenced might have suffered some negatives at levels. Combining alternative methods may therefore more reliable conclusions pathogen's presence. Further, we found correlation between prevalence infection carriers in average amounts detected infected local individuals.