FIELD: biotechnology.
SUBSTANCE: invention relates to medical biotechnology and describes a method for detecting parvovirus B19 at low viral load by detecting virus DNA in a blood sample by PCR with further fragment sequencing. Detection is carried out in plasma and / or serum, oropharyngeal washes. Diagnostics is carried out in two stages. At the first stage, asymmetric PCR with extended primers is used (forward primer 5'-agccattttaagtgttttactataattttattggtcagtttt-3', reverse primer 5'-tgttgttcatatctggttaaatacttaaattc-3'). At the second stage, primers are used to regions of the parvovirus B19 genome, while there is a uniform ratio of deoxynucleoside triphosphates with a reduced concentration and a high concentration of MgCl2 in the composition of the amplification mixture at each stage; at the first stage, the mixture contains glycerol in an amount of 5% of the final volume, and at the second stage it contains formamide and DMSO in the amount of 4% and 8% of the final volume, respectively.
EFFECT: method increases sensitivity and specificity of the diagnosis.
1 cl, 2 dwg, 2 tbl, 3 ex
