EMILIO AGUINALDO COLLEGE

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SCHOOL OF MEDICAL TECHNOLOGY

Ricafort, Sean Christopher R. IS Lecture

MMLS 3-2

Journal Critiquing

Final Period Topic: Diagnosis of Prions (Molecular, Serological)

Title: Ultrasensitive RT-QuIC Seed Amplification Assays for Disease-Associated Tau, α-

Synuclein, and Prion Aggregates

1. Relevance of the objective to the study

- The objective of this research is to provide a new method for identifying protein
aggregates such as prions. The goal is important because there is no known treatment
for deadly prion disorders. Early detection and treatment of these illnesses could
result from a new method for detecting prions, which could improve patient
outcomes. This is also important since it might help with the creation of novel prion
disease treatments. RT-QuIC can be used to track the success of prion disease
treatments. The usage of this data helps optimize treatment plans and enhance
patient outcomes.
2. Short summary of the methodology
- The K19CFh protein is expressed in the BL21 (DE3) strain of Escherichia coli using the
Overnight Express autoinduction method. After confirming the expression of the
target gene and obtaining a good yield of the recombinant protein, the cells are
stored in glycerol at -80 °C. The protocol involves plating cells from the frozen
bacterial glycerol stock, inoculating a starter culture, and growing it at 37 °C with
shaking. An overnight autoinduction culture is then prepared, followed by the
preparation of cell extracts using FPLC chromatography. The protein is purified using
a nickel column, and the fractions containing K19CFh are collected. Acetone
precipitation and desalting steps are performed to further purify the protein, and the
purified K19CFh is then aliquoted, frozen, and stored. Another round of purification is
performed using the second filtrate from the cell extracts. The method also includes
the preparation of tau RT-QuIC reactions for protein analysis and provides
instructions for data analysis.
3. Short summary of the results, discussion, and conclusion
- According to the result of the study, the RT-QuIC assay was successful in identifying
disease-related tau, -synuclein, and prion aggregates in every sample examined. It is
possible that the assay is a very sensitive method for detecting protein aggregates
because it was able to detect these aggregates at extremely low concentrations.
Alzheimer's patients' cerebrospinal fluid was found to contain disease-related tau
clumps by the RT-QuIC assay. In the brain tissue of Parkinson's disease patients, the

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Gov. D. Mangubat Ave., Brgy. Burol Main, City of Dasmariñas, Cavite 4114, Philippines
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SCHOOL OF MEDICAL TECHNOLOGY

assay was also able to find disease-related -synuclein aggregates. Last but not least,
the RT-QuIC assay proved successful in identifying disease-associated prion
aggregates in skin tissue from Creutzfeldt-Jakob disease patients. The result suggests
that the RT-QuIC test can be a useful tool for the early detection of
neurodegenerative disorders. The assay could be used to determine which patients
are more likely to contract these illnesses, as well as to track the disorders'
development. Therefore, this method can be used for the early diagnosis of
neurodegenerative diseases. The RT-QuIC assay is still in its early stages of
development, but it has the potential to revolutionize the way we diagnose and treat
neurodegenerative diseases.
4. Limitations and possible suggestions to the journal article
- The study contains some kind of limits. First off, the study only used a few samples,
therefore it's crucial to confirm the results in a broader investigation. Second,
because the study was cross-sectional in nature, the researchers were unable to
ascertain what caused the association between protein aggregation and
neurodegenerative disorders. Third, the clinical relevance of the discovered protein
aggregates was not evaluated by the study. It's likely that the protein aggregates that
were found have nothing to do with symptoms of disease. With this limitations,
several suggestions can be done for future research related to the study which
includes conducting a larger study with a more diverse population of patients, using a
longitudinal design to track the progression of protein aggregates over time,
Assessing the clinical significance of protein aggregates by correlating their levels with
clinical symptoms, and developing a treatment that targets protein aggregates.
5. What is the societal impact of the journal article?
- The RT-QuIC assay could also lead to the development of new treatments for
neurodegenerative diseases. If it is possible to target protein aggregates, this could
lead to the development of drugs that can prevent or slow the progression of these
diseases. This could have a major impact on the lives of millions of people who are
affected by neurodegenerative diseases. Researchers can lead the development by
gathering more information through large quantity tests to see if it is totally effective.

Reference:

Saijo, E. (2018). Ultrasensitive RT-QuIC Seed Amplification Assays for Disease-Associated Tau, α-

Synuclein, and Prion Aggregates Protein Misfolding Diseases, 19–37. Doi: 10.1007/978-1-

4939-8820-4_2

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