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Related Experiment Video

Updated: Oct 6, 2025

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Sensitive, Single-Pot, Two-Stage Assay for Hepatitis Viruses.

Youngung Seok1, Qingtian Yin1, Huiwen Bai1

  • 1Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania (UPenn), 216 Towne Building, 220 South 33rd Street, Philadelphia, Pennsylvania 19104, United States.

Analytical Chemistry
|January 13, 2022
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Summary

New point-of-care tests for hepatitis B (HBV) and hepatitis C (HCV) enable rapid diagnosis and treatment. The sensitive Penn-RAMP assay provides results in 30 minutes, improving patient follow-up and care.

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Area of Science:

  • Molecular biology
  • Infectious diseases
  • Point-of-care diagnostics

Background:

  • Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections can lead to severe illness if untreated.
  • Asymptomatic nature of these infections necessitates routine screening for at-risk populations.
  • Current diagnostic methods involve multiple clinic visits, leading to poor patient follow-up.

Purpose of the Study:

  • To develop sensitive, rapid, point-of-care tests for HBV and HCV detection.
  • To enable a "test and treat" strategy for hepatitis B and C.
  • To overcome the limitations of current multi-step diagnostic procedures.

Main Methods:

  • Development of a two-stage, isothermal molecular amplification assay named Penn-RAMP.
  • Utilizing recombinase polymerase amplification (RPA) in the first stage and loop-mediated isothermal amplification (LAMP) in the second stage.
  • Implementing a single-pot system with a thermally removable barrier for sequential amplification.

Main Results:

  • Penn-RAMP demonstrated higher sensitivity compared to RPA or LAMP alone.
  • The assay successfully detected as few as 10 HBV virions and 25 HCV virions within 30 minutes.
  • Viral load estimation is possible based on signal threshold time.

Conclusions:

  • Penn-RAMP offers a sensitive and rapid point-of-care solution for HBV and HCV detection.
  • This technology facilitates a "test and treat" approach, potentially improving patient outcomes.
  • The assay's design allows for minimal instrumentation and efficient workflow.