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Technical Notes
Technology Note
Multiplex unleashed with Hyperplex PCR™: Validating the 100-Plex Breakthrough
In this technical note, we delve into the technical performance of hpPCR™, showcasing its sensitivity beyond digital PCR and single-nucleotide specificity out of the box, while introducing a new paradigm of multiplex.
Publications
Simultaneous detection of RNA and DNA from pathogens with Hyperplex PCR
Sofie Bergstrand, Linnea Blom, Paula Ågren, Johan Aarum, Johannes Hedman
This study presents the first application of hpPCR for simultaneous detection of RNA and DNA from pathogens, using a 16-plex panel for gastrointestinal pathogens relevant in food and waterborne outbreaks. The panel showed no false positives or cross reactions and detection limits comparable to single-plex qPCR and RT-qPCR.
Hyperplex PCR for forensic mRNA-based cell typing
Maja Sidstedt, Margreet van den Berge, Titia Sijen, Johannes Hedman
This study evaluates hpPCR for forensic mRNA-based cell type identification, demonstrating equivalent or enhanced detection compared with the CE-based CellTyper method across two forensic laboratories, including improved sensitivity for minority contributions in mixed samples.
Integration of periodontal pathogens and inflammatory mediators in saliva as biomarkers for periodontitis
Carl Titusson, Anna Lundmark, Ruben R. G. Soares, Anastasios Damdimopoulos, Gunnar Johannsen, Umear Naseem, Tülay Yucel-Lindberg
This study used hpPCR to profile periodontal pathogens in saliva from 57 participants, identifying Fretibacterium spp. as the strongest discriminator between periodontal health and disease (AUC = 0.82), which served as a step toward non-invasive, saliva-based periodontitis diagnostics.
20-Plex Gene Expression Analysis Using Hyperplex PCR (hpPCR) Toward the Development of Salt-Tolerant Wheat
Esteri Viitanen, Samuel Narmack, Margarita Psallida, Johan Banér, Umear Naseem, Ruben R. G. Soares, and Henrik Aronsson
This chapter presents a 20-plex hpPCR gene-expression assay applied to uncover transcripts associated with salt tolerance in wheat.
Hyperplex PCR enables highly multiplexed analysis of point mutations in wastewater: Long-term SARS-CoV-2 variant surveillance in Sweden as a case study
Ruben R.G. Soares, Javier Edo Varg, Attila Szabó, Mariana Kluge, Filip Petrini, Margarita Psallida, Paweł Olszewski, Danai V. Nikou, Isaac Owusu-Agyeman, Mariel Perez-Zabaleta, Zeynep Cetecioglu, Umear Naseem, Maja Malmberg, Anna J. Székely
The publication, produced in collaboration with SEEC, showcases a full year of SARS-CoV-2 variant tracking in Swedish wastewater using hpPCR. This study demonstrates how hpPCR successfully fills the bioanalytical method gap between qPCR/dPCR and NGS, upgrading the current WBE analytical toolbox.
Long-term SARS-CoV-2 surveillance in the wastewater of Stockholm: What lessons can be learned from the Swedish perspective?
Mariel Perez-Zabaleta, Amena Archer, Kasra Khatami, Mohammed Hakim Jafferali, Prachi Nandy, Merve Atasoy, Madeleine Birgersson, Cecilia Williams, Zeynep Cetecioglu
This study presents the learning outcomes from over two-year long monitoring of SARS-CoV-2 in Stockholm, Sweden, where hpPCR was used as proof-of-concept for variant profiling.
Rolling Circle Amplification in Integrated Microsystems: An Uncut Gem toward Massively Multiplexed Pathogen Diagnostics and Genotyping
Ruben R. G. Soares, Narayanan Madaboosi, and Mats Nilsson
A review introducing padlock probes, RCA and framing their untapped potential towards unprecedented multiplexing (hyperplexing) when powered by APLEX Bio's Nanopixels™