Evaluation of Real Time PCR measurement rhizopus oryzae in tomato paste

In this study, Molecular method to measure the rhizopus oryzae Real Time PCR was used in tomato paste. Because of problems mold count HMC method such as Proofer error, low and non-repeatability precision, accuracy and repeatability developing a standard method that can enable high accuracy and repeatability and mold counts it seems necessary. Real Time PCR molecular methods can be used as a suitable method to identify and quantify the precise mold is considered that based on the measurement of absolute and relative genes is the existing molds. Tomato paste samples with different amounts of mold spores (101, 103 and 105) were contaminated per gram. After incubation and growth of mold spores and mycelium extraction DNA from each sample was performed using al-sammari method. Then, using Real Time PCR reaction of DNA copy number of molds using SYBR Green reagent and primers of the rhizopus oryzae genus was determined. The results showed that increasing amount of DNA molds in controls and inoculated samples resulted Real Time curve in the lower CT values are entered logarithmic phase. The correlation coefficient of linear calibration Real Time was R2 = 0/943. This indicate that the accuracy of this test is quantitative detection. The reaction specificity was determined using melting curve of the primers and indicated that the reaction has been completely dedicated.