Multiplex Pathogen Sequencing
Multiplex Pathogen Sequencing - Related Videos
03:45
A Bead-Based Multiplex Immunoassay to Identify Pathogen-Specific Antibodies in Saliva
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2025
This video demonstrates a bead-based multiplex immunoassay technique to simultaneously detect antibodies for multiple environmental pathogens in human saliva. Fluorescent beads were coupled to antigens from different pathogens, which were captured using antibodies in the human saliva sample. Upon labeling the antibodies using a fluorescent reporter, the beads were analyzed using flow cytometry to assess the presence of different pathogen-specific antibodies in the saliva.
09:08
Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens
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Cited by 11 •
2016
In the current climate of scarce resources, new technologies are emerging that allow researchers to conduct studies cheaper, faster and with more precision. Here we describe the development of a bead-based salivary antibody multiplex immunoassay to measure human exposure to multiple environmental pathogens simultaneously.
06:34
Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
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Cited by 29 •
2016
Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and molecular techniques (16S rRNA gene sequencing) permit the identification of rare bacterial pathogens in routine diagnostics. The goal of this protocol lies in the combination of both techniques which leads to more accurate and reliable data.
09:54
Postmortem Diagnosis of Rabies in Animals by the Updated, Multiplexed LN34 Real-Time Reverse Transcription-Polymerase Chain Reaction Assay
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Cited by 2 •
2025
This protocol demonstrates the pan-lyssavirus LN34 real-time reverse transcription-polymerase chain reaction (RT-PCR) assay from tissue collection to result interpretation, including updates to primer sequences and formulations to improve assay performance for some non-rabies lyssaviruses and lagomorphs. We also demonstrate the assay setup for a single-well LN34 multiplexed (LN34M) format.
08:30
Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells
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Cited by 4 •
2020
Here we presented a multiplexed single cell mRNA sequencing method to profile gene expression in mouse embryonic tissues. The droplet-based single cell mRNA sequencing (scRNA-Seq) method in combination with multiplexing strategies can profile single cells from multiple samples simultaneously, which significantly reduces reagent costs and minimizes experimental batch effects.
FAQs
Sample-specific barcodes distinguish material from different specimens after they enter a shared sequencing workflow. Once sequencing is complete, bioinformatic analysis can associate reads with the appropriate sample before comparing genetic material with reference genomes. This organization allows several specimens or pathogen targets to be processed together while preserving the identity needed for clinical interpretation.
Reference genomes provide the comparison framework for identifying pathogen genetic material and recognizing differences among closely related strains. Bioinformatic comparison can therefore move beyond simple detection and support characterization of the organisms present. In medicine, this distinction may clarify which infectious agents are represented when clinical symptoms alone do not identify the cause.
Reliable interpretation depends on careful validation because sequencing may detect contamination or background reads in addition to clinically relevant material. Validation helps establish which signals are credible and how findings should be interpreted. This safeguard is especially important when several pathogens are assessed in one workflow, since weak or unexpected signals could otherwise be mistaken for evidence of infection.
Analyzing multiple genetic targets in the same specimen can reveal more than one infectious agent, helping identify mixed infections that a single-agent result could miss. Genetic comparison also supports separation of closely related strains. These capabilities can improve clinical characterization when nonspecific symptoms do not clearly indicate whether one pathogen, several pathogens, or a particular strain is involved.
The workflow begins with nucleic-acid extraction from a clinical specimen, followed by either targeted amplification or library preparation. Sample-specific barcodes may then organize material for next-generation sequencing. Finally, bioinformatic comparison with reference genomes identifies and characterizes the genetic material detected. Each stage contributes to turning a mixed molecular signal into an interpretable result.
The approach may be valuable when symptoms are nonspecific and routine clinical assessment does not clearly identify the infectious cause. Testing several pathogens within one workflow can also conserve limited clinical specimens and may reduce testing time compared with separate investigations. Results can provide broader diagnostic information while still requiring validation before clinical interpretation.
Findings may help guide infection-control decisions by identifying the pathogen or strain present in a clinical specimen. At a broader level, repeated sequencing results can support surveillance of emerging variants and antimicrobial-resistance markers. This connects individual diagnostic testing with public-health monitoring, allowing genetic changes and clinically important resistance-related signals to be tracked.