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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

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Real-time Quaking-induced Conversion Assay for Detection of CWD Prions in Fecal Material
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Real-time quaking-induced conversion: a highly sensitive assay for prion detection.

Ryuichiro Atarashi1, Kazunori Sano, Katsuya Satoh

  • 1Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. atarashi@nagasaki-u.ac.jp

Prion
|July 23, 2011
PubMed
Summary

A new real-time quaking-induced conversion (RT-QUIC) assay detects abnormal prion protein (PrPSc) in cerebrospinal fluid with high accuracy. This sensitive and specific test aids in the early diagnosis of prion diseases like Creutzfeldt-Jakob disease.

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Infectious Diseases

Background:

  • Prion diseases are fatal neurodegenerative disorders.
  • Early and accurate diagnosis is crucial for patient management.
  • Current diagnostic methods for prion diseases have limitations.

Purpose of the Study:

  • To develop and validate a novel in vitro amplification technology for prion detection.
  • To assess the diagnostic accuracy of the developed assay for sporadic Creutzfeldt-Jakob disease (sCJD).
  • To evaluate the applicability of the assay to various prion diseases and its quantification capabilities.

Main Methods:

  • Development of real-time quaking-induced conversion (RT-QUIC) assay.
  • Testing of over 200 cerebrospinal fluid (CSF) specimens from Japanese and Australian patients.
  • Application of RT-QUIC to samples from scrapie, chronic wasting disease (CWD), and bovine spongiform encephalopathy (BSE).
  • Quantification of prion seeding activity using end-point dilution.

Main Results:

  • The RT-QUIC assay demonstrated no false positives in over 200 CSF specimens.
  • Achieved over 80% accuracy in diagnosing sporadic Creutzfeldt-Jakob disease (sCJD).
  • Successfully applied to detect prions in scrapie, CWD, and BSE.
  • Enabled quantification of prion seeding activity.

Conclusions:

  • RT-QUIC is a highly sensitive and specific in vitro diagnostic assay for prion diseases.
  • The assay facilitates early, rapid, and accurate diagnosis of sCJD.
  • RT-QUIC shows broad applicability across different prion diseases and offers quantification of prion load.