A new study in Analytical Biochemistry presents the first application of Hyperplex PCR™ to the simultaneous detection of RNA and DNA from pathogens.

Researchers from the National Forensic Centre of the Swedish Police Authority, Lund University, the Swedish Food Agency and the Public Health Agency of Sweden developed and evaluated a 16-plex panel for gastrointestinal pathogens relevant in food and waterborne disease outbreaks. The panel covers RNA viruses (Hepatitis A, Norovirus GI and GII), a parasite (Cryptosporidium parvum) and bacteria, including highly pathogenic species such as Bacillus anthracis and Yersinia pestis. An initial reverse transcription step allows RNA and DNA targets to be detected in the same reaction.

Key results:

• Selectivity: all expected targets were detected, with no false positives or cross reactions across the 11 bacteria, 1 parasite and 4 viruses tested.

• Sensitivity: detection limits were comparable to single-plex qPCR and RT-qPCR assays using the same primers.

• Inhibitor tolerance: hpPCR tolerated high amounts of humic acid, a model inhibitor for food and environmental samples.

• Room to grow: the panel can be supplemented with further assays, as hpPCR supports up to 100 targets in one reaction.
  

The authors position hpPCR as an alternative or complement to qPCR and sequencing for rapid, multiplex detection of pathogens in food and environmental samples with unknown contents.

Read the paper (open access): Bergstrand S, Blom L, Ågren P, Aarum J, Hedman J. Simultaneous detection of RNA and DNA from pathogens with hyperplex PCR. Analytical Biochemistry 720 (2027) 116266. https://doi.org/10.1016/j.ab.2026.116266