Bashkir Medical University, Ufa; Gamaleya Research Institute of Epidemiology and Microbiology, Moscow, Russia
The information on the applied aspects of the polymerase chain reaction (PCR) is updated. In particular, the main inhibitor of PCR, considerably decreasing the sensitivity of the method both at the lysis stage of the tested material and due to the degradation of the DNA matrix and primers and/or to the direct inhibition of the activity of DNA polymerase, are described. The compounds, most frequently distorting the course of the reaction while testing clinical blood samples, bioptic samples, sputum, etc., are characterized. Testing concrete clinical material with the use of PCR was shown to require differentiated approach both at the stage of choosing the adequate method for the preparation of samples and at all other stages, including, e.g., the corresponding DNA polymerases or at the stage of heating for decreasing endonuclease activity or for IgG denaturation. Information on the causes of false negative results of PCR and the variants of their elimination, useful under practical laboratory conditions, is given.
Zh.Mikrobiol. (Moscow), 2003, No. 3, P. 93—98