In a new study, researchers developed and evaluated a novel hexaplex PCR assay for simultaneous detection of pathogenic B. anthracis and Y. pestis as well as discrimination with non-pathogenic strains in a single PCR tube.

A multiplex PCR assay comprising a total of six primer pairs for B. anthracis and Y. pestis detection, respectively, was developed.

In the assay, the recombinant internal amplification control (IAC) was designed to control both DNA extraction and PCR processing, allowing the evaluation of the true negative to false negative results. The researchers used no additional adjuvant supplementary materials, which was recommended by various authors to improve amplification efficiency and specificity of the multiplex PCR.

Thirty-eight bacterial strains including 4 reference strains and 34 screening strains derived from human, herbivores, and environments in the North of Vietnam, as well as spiked soil samples, were used to evaluate the assay.