METHOD FOR DETERMINATION OF BIOMATERIALS BACTERIAL CONTAMINATION Russian patent published in 2017 - IPC C12Q1/00 

FIELD: biotechnology.

SUBSTANCE: method for determining bacterial contamination of human cell cultures, as well as medicine and biomaterials based on them via PCR-amplification of the DNA sequence of the bacterial 16s RNA gene, universal for all bacterial species, differs in that it has internal control, is free from false positive results and controlled sensitivity, which is achieved by introducing primers into the reaction mixture to amplify the sequence of the human beta-actin gene. The method provides control of DNA purification from the sample and the PCR passage; moreover, the reference reaction competes with the amplification reaction of the 16s RNA gene, as a result of which the amplification of small amounts of bacterial DNA presenting in the reaction mixture components is suppressed.

EFFECT: change in the temperature of primers annealing-elongation in this method results in the change in the reference reaction efficiency, which enables to regulate the sensitivity threshold of the proposed method.

3 cl, 2 dwg, 3 ex