Determination of cycle cut off in real‐time PCR for the detection of regulated plant pathogens

Abstract

Given the high importance of an accurate detection for quarantine plant pathogens, and the risk of false positive or false negative results, a detection method must be correctly validated. A statistical procedure has been developed to determine the cycle cut off and the corresponding limit of detection in real‐time PCR, considering a risk of false positive results of 1% and a probability of detection of 95%. The procedure presented in this paper is easy to develop and can be applied to any plant pathogen to be detected by real‐time PCR.