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High capacity and low cost detection of prion protein gene variant alleles by denaturing HPLC
Castro RM, Landemberger MC, Walz R, Carlotti CG, Huang N, Cunha DR, Moura R, Caballero OL et al.. Journal of neuroscience methods, 2004-10-01
Topic: Brain proteins, microglia and lab models
Mutations in the human prion protein gene (PRNP) are responsible for hereditary diseases called transmissible spongiform encephalopathies (TSE) and a polymorphic site at codon 129 determines sensitivity to infectious forms of these maladies. More recently, codon 129 has been related to cognition performance in the elderly, in Alzheimer disease (AD) and in Down syndrome. Furthermore, a rare polymorphism at codon 171 was described in 23% of patients with mesial temporal lobe epilepsy related to hippocampal sclerosis (MTLE-HS), the most common form of surgically remediable epileptic syndrome. Thus, a method that permits fast and efficient screening of PRNP mutations and polymorphisms in patients, in high risk populations, and in family members is desirable. In the present study, we established the conditions for analysis of the PRNP open reading frame using denaturing high-performance liquid chromatography (DHPLC), whereby unpurified PCR products were subjected to denaturing and reannealing steps leading to heteroduplex formation. We described specific profiles for the PRNP polymorphisms at codons 129 (M/V), 117 (A/A silent), 219 (E/K), 171 (N/S), and the octarepeat deletion using amplified DNA from 562 samples. The chromatograms for TSE-associated mutations at codons 102 (P/L), 183 (T/A), and 210 (V/I) were also determined. Specificity of the DHPLC profile for each PRNP variant allele was confirmed in 100% of the samples by direct and cloned DNA sequencing in addition to endonuclease digestion when applicable. Therefore, the present study shows that DHPLC is a rapid, highly accurate and efficient technique for the detection of PRNP genetic variants.
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APA 7th
Castro, R. M., Landemberger, M. C., Walz, R., Carlotti, C. G., Huang, N., Cunha, D. R., et al. (2004). High capacity and low cost detection of prion protein gene variant alleles by denaturing HPLC. Journal of neuroscience methods. https://doi.org/10.1016/j.jneumeth.2004.05.001
Vancouver
Castro RM, Landemberger MC, Walz R, Carlotti CG, Huang N, Cunha DR, et al. High capacity and low cost detection of prion protein gene variant alleles by denaturing HPLC. Journal of neuroscience methods. 2004. doi:10.1016/j.jneumeth.2004.05.001. PMID: 15488240.
BibTeX
@article{castro2004-15488240,
author = {Castro, R. M. and Landemberger, M. C. and Walz, R. and Carlotti, C. G. and Huang, N. and Cunha, D. R. and Moura, R. and Caballero, O. L. and others},
title = {{High capacity and low cost detection of prion protein gene variant alleles by denaturing HPLC}},
journal = {Journal of neuroscience methods},
year = {2004},
doi = {10.1016/j.jneumeth.2004.05.001},
pmid = {15488240},
url = {https://doi.org/10.1016/j.jneumeth.2004.05.001}
}
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Key findings
- In the present study, we established the conditions for analysis of the PRNP open reading frame using denaturing high-performance liquid chromatography (DHPLC), whereby unpurified PCR products were subjected to denaturing and reannealing steps leading to heteroduplex formation.
- We described specific profiles for the PRNP polymorphisms at codons 129 (M/V), 117 (A/A silent), 219 (E/K), 171 (N/S), and the octarepeat deletion using amplified DNA from 562 samples.
- The chromatograms for TSE-associated mutations at codons 102 (P/L), 183 (T/A), and 210 (V/I) were also determined.
- Specificity of the DHPLC profile for each PRNP variant allele was confirmed in 100% of the samples by direct and cloned DNA sequencing in addition to endonuclease digestion when applicable.
- Therefore, the present study shows that DHPLC is a rapid, highly accurate and efficient technique for the detection of PRNP genetic variants.
Why it matters
This study established conditions for detecting prion protein gene variants, including codon 129, using denaturing HPLC. It is a laboratory method for genetic screening and reports no treatment, cure or prevention for Alzheimer's disease.
Study details
- Evidence strengthLow
How this study scored 31/100
Published in a peer-reviewed journal. Scored zero for study design, sample size, effect reported, studied in people and randomized / blinded because the record does not state it.
| Component | Points | Why |
|---|---|---|
| Journal tier | 7 / 10 | Tier 3 — peer-reviewed journal (Journal of neuroscience methods) |
| Journal's citation record | 2 / 4 | Journal of Neuroscience Methods: journal h-index 227 (OpenAlex, 2026-10-29) |
| Study design | 0 / 12 | No study design recorded — scores zero, not guessed (not stated in the record — scored 0) |
| Strength of evidence | 1 / 5 | Rated “low” in our summary |
| Peer-review validation | 10 / 10 | Peer-reviewed. Published as an article in a named peer-reviewed journal. |
| How recent | 1 / 5 | 2004 — older than 20 years, still counted |
| Sample size | 0 / 4 | Participant count not stated — scores zero (not stated in the record — scored 0) |
| Effect reported | 0 / 3 | No effect size reported — scores zero (not stated in the record — scored 0) |
| Studied in people | 0 / 3 | Not stated whether this was studied in people — scores zero (not stated in the record — scored 0) |
| Randomized / blinded | 0 / 2 | No randomization or blinding stated — scores zero (not stated in the record — scored 0) |
| Times cited | 2 / 7 | Cited 11 times (OpenAlex, 2026-10-10) |
| Citations per year | 0 / 5 | 0.5 citations a year over 24 year(s) since 2004 |
| Impact for its field and year | 1 / 6 | Cited 0.3× the average for papers of its field and year (OpenAlex FWCI) |
| Cited by studies here | 0 / 6 | No other study in this library cites it |
| Still being cited | 0 / 4 | Cited 0 times in 2024–2026 (2026 still in progress; OpenAlex) |
| Senior author's track record | 2 / 4 | Last-listed author Vilma Regina Martins: h-index 49 (OpenAlex, whole career) |
| Institution's track record | 1 / 3 | Universidade de São Paulo: institution h-index 936 (OpenAlex) |
| Multicenter collaboration | 2 / 3 | OpenAlex lists 4 institution(s) in 4 countries |
| Indexed in PubMed | 2 / 2 | Indexed in PubMed (PMID 15488240) |
| Free to read | 0 / 2 | Behind a paywall — scores zero |
| Total | 31 / 100 | — |
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