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Related Concept Videos

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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Related Experiment Video

Updated: Apr 14, 2026

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
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Focused human gene expression profiling using dual-color reverse transcriptase multiplex ligation-dependent probe

Mariëlle C Haks1, Jelle J Goeman2, Cecile Magis-Escurra1

  • 1Department of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.

Vaccine
|April 29, 2015
PubMed
Summary
This summary is machine-generated.

We developed a dual-color Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification (dcRT-MLPA) assay to rapidly profile mRNA expression for studying human immune responses. This high-throughput method identifies predictive host biomarker signatures in large human cohorts.

Keywords:
Gene expression profilingHost biomarkersdcRT-MLPA

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genomics

Background:

  • Investigating human immune responses to vaccines or infections requires scalable methods for profiling host gene expression.
  • Existing techniques may have limitations in throughput or dynamic range for large-scale cohort studies.

Purpose of the Study:

  • To introduce and validate a novel dual-color Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification (dcRT-MLPA) assay.
  • To demonstrate the utility of dcRT-MLPA for high-throughput host biomarker profiling in human population studies.

Main Methods:

  • Development of a dual-color Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification (dcRT-MLPA) assay.
  • Application of lasso regression analysis and RUV-2 for identifying host biomarker signatures.
  • Profiling mRNA expression of multiple host genes from various human tissues and body fluids.

Main Results:

  • dcRT-MLPA offers dynamic range and sensitivity comparable to real-time QPCR and RNA-Sequencing.
  • The assay is suitable for high-throughput monitoring of host biomarkers in large human cohorts (cross-sectional and longitudinal).
  • Identified multicomponent host biomarker signatures with excellent predictive values.

Conclusions:

  • dcRT-MLPA is a versatile and powerful platform for investigating human immune responses at a population scale.
  • The assay facilitates the discovery of predictive host biomarker signatures and comparison across diverse populations.
  • dcRT-MLPA can be adapted for various tissues and body fluids, enhancing its applicability.