US 12,265,089 B2 · Discriminating Parkinson's disease from multiple system atrophy using alpha-synuclein PMCA

Abstract

A method is provided for distinguishing between and/or diagnosing Parkinson's disease (PD) or multiple system atrophy (MSA) in a subject who is exhibiting symptoms associated with both PD and MSA. The method comprises: (A) contacting a biological sample obtained from the subject and comprising soluble, misfolded alpha-synuclein (αS) protein with a pre-incubation mixture comprising a monomeric αS substrate and an indicator to form an incubation mixture; (B) conducting an incubation cycle two or more times on the incubation mixture to form misfolded αS aggregates; (C) subjecting the incubation mixture to excitation and detecting via indicator fluorescence emission the misfolded αS aggregates; and (D) diagnosing the subject has having PD or MSA depending on the fluorescence emission intensity. In some aspects, the incubation cycles are conducted in the presence of a bead.

Claims43

Dependent claims

A method for differentiating a diagnosis of Parkinson's Disease (PD) from a diagnosis of Multiple System Atrophy (MSA) in a human subject, the method comprising:

(A) contacting a cerebrospinal fluid (CSF) or a brain homogenate (BH) sample (i) obtained from the human subject and (ii) comprising soluble, misfolded alpha-synuclein (αS) protein, with a pre-incubation mixture, the pre-incubation mixture comprising:
(1) a monomeric αS substrate;
(2) a buffer composition;
(3) sodium chloride (NaCl);
and
(4) a fluorescent protein aggregation indicator,
to form an incubation mixture;
(B) incubating and agitating the incubation mixture to form an incubated mixture comprising misfolded αS aggregates;
(C) illuminating the incubated mixture with a wavelength of light that excites the fluorescent protein aggregation indicator, including:
(1) determining an initial average fluorescence (AFi) of the incubated mixture prior to the formation of the misfolded αS aggregates;
(2) determining a standard deviation (SDi) of the AFi;
and
(3) determining a maximum fluorescence (Fmax) of the incubated mixture after the formation of the misfolded αS aggregates;
and
(D) diagnosing the human subject as having PD or MSA, where:
(1) if the Fmax is from about 22.2×SDi+AFi to 250×SDi+AFi, the patient is diagnosed as having MSA;
and
(2) if the Fmax is above 250×SDi+AFi, the patient is diagnosed as having PD.

15 more elements

Dependencies are inferred from the claim text.

Discriminating Parkinson's disease from multiple system atrophy using alpha-synuclein PMCA

Life of this patent

Discriminating Parkinson's disease from multiple system atrophy using alpha-synuclein PMCA

Life of this patent

Abstract

Claims43