Evaluation of correlation between quality and amount of the extracted viral RNA from raw milk with the amount of raw milk dry Matter for molecular detection of enteric viruses using response surface model

Nowadays, presence of enteric viruses in water and some foods is proved as a cause for infectious diseases. Raw milk can transmit enteric viruses potentially. Lack of distinct protocol for recovery and extraction of these viruses from raw milk made us to evaluate the effect of dry matter amount on quality and amount of viral RNA yield. Therefore according to reverse of routine protocols in separating of milk components, we prepared four milk model solutions in different combinations and ratios based on natural raw milk components. Then, each milk model solution was spiked with five levels of MS2 coliphage (1.3×10⁴, 1.3 ×10², 1.3, 1.3×10^-2, 1.3 ×10^-4 pfu/ml). We isolated the viral RNA from each milk model solution using phenol - guanidine thiocyanat - chloroform method. Also we measured quality and RNA extraction Yield in twelve frequencies by nanodrop spectrophotometer. The results showed that in the equal viral concentration, the most amount of viral RNA was extracted from whole milk model solution (including fat). Whereas casein and whey proteins had the most inhibition in the viral RNA extraction .Also the highest quality of the extracted RNA for molecular detection was related to whole milk model solution. Based on response surface model (polynomial) RNA extraction yield and quality of the extracted RNA were closely correlated to the amount of dry matter of milk model together with coliphage concentration (R=0/96 and R= 0/92 respectively).