Nonaka, T.; Iwasaki, Y.; Horiuchi, H.; Satoh, K. Detection Limitations of Prion Seeding Activities in Blood Samples from Patients with Sporadic Prion Disease. BMC Neurology 2024, 24, doi:10.1186/s12883-024-03590-7.
Nonaka, T.; Iwasaki, Y.; Horiuchi, H.; Satoh, K. Detection Limitations of Prion Seeding Activities in Blood Samples from Patients with Sporadic Prion Disease. BMC Neurology 2024, 24, doi:10.1186/s12883-024-03590-7.
Nonaka, T.; Iwasaki, Y.; Horiuchi, H.; Satoh, K. Detection Limitations of Prion Seeding Activities in Blood Samples from Patients with Sporadic Prion Disease. BMC Neurology 2024, 24, doi:10.1186/s12883-024-03590-7.
Nonaka, T.; Iwasaki, Y.; Horiuchi, H.; Satoh, K. Detection Limitations of Prion Seeding Activities in Blood Samples from Patients with Sporadic Prion Disease. BMC Neurology 2024, 24, doi:10.1186/s12883-024-03590-7.
Abstract
Human prion diseases (HPDs) are fatal neurodegenerative disorders characterized by abnormal prion proteins (PrPSc). Numerous techniques have been devised to detect prion seeding activity, each with limitations. Here, we developed a technology for in vitro amplification of abnormal prion proteins (PrPSc), thereby enabling their sensitive detection. Using the real-time quaking-induced conversion (RT-QuIC) PrPSc amplification assay, we could detect ≥1 fg of PrPSc in diluted human prion disease brain homogenate. Although RT-QuIC assay is suitable for detecting prion seeding activity in a variety of specimens, its accuracy in whole blood, blood plasma, or blood-contaminated tissues is reduced. To circumvent this issue and move toward developing a blood test for prions, we introduced an immunoprecipitation step at the beginning of the RT-QuIC reaction. We refer to this enhanced method as “eQuIC”. eQuIC utilizes three types of human monoclonal antibodies, which significantly increased its sensitivity—making it approximately 1000 times more sensitive than the original RT-QuIC assay. Nevertheless, when we tested the blood (serum, plasma, and whole blood) of six patients with sporadic human prion disease using eQuIC, we were unable to detect prion activity. Therefore, we concluded that the detection of prion seeding activities in blood samples from patients with sporadic prion disease remains challenging; thus, alternative methods beyond RT-QuIC and eQuIC will be necessary for future research.
Medicine and Pharmacology, Neuroscience and Neurology
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