[Comparative analysis of LAMP and Real Time PCR methods to detect pathogens of glanders and meliodosis.]

Results of detection of Burkholderia mallei and Burkholderia pseudomallei DNA strains by LAMP (Loop-mediated Isothermal Amplification) and Real Time PCR are shown. It has been revealed that, in Real Time PCR, primers steadily detected DNA of those microorganism for the sequences of which they were designed. The above mentioned primers did not detect DNA of heterologous strains. During LAMP method no set of primers showed high analytical sensitivity and specificity. Primers did not detected DNA of all the strains under research to target genes of which they were not intended, but they were capable of directing the synthesis of fragments of genes of heterologous strains. Furthermore, it was difficult to reach the same results during repeated experiments. Failures during LAMP may occur due to existence of GC-reach regions in Burkholderia mallei and Burkholderia pseudomallei genomes and due to emergence of secondary structures in isothermical conditions. It is recommended to use Real Time PCR in order to detect pathogens, in case of such matrixes as Burkholderia mallei and Burkholderia pseudomallei DNAs which are very complicated for LAMP.