Granted Patent B2

US 12,467,094 · App. 17/692,408 · Granted Nov 11, 2025

Inventor: Derek Croote (South San Francisco, CA)

Assignee: IGGENIX, INC.

C12Q1/6883C12Q1/6851C12Q2600/118C12Q2600/158

Abstract

Claims (23)

1 . A method for predicting a future allergic response to one or more allergens in a subject, the method comprising:

obtaining a blood sample from the subject;

enriching B cells from the blood sample;

lysing the enriched B cells to thereby release RNA;

measuring and identifying one or more RNA species comprising a nonproductive epsilon germline transcript (&GLT) in the blood sample by performing reverse transcription followed by PCR to produce cDNA from the released RNA,

wherein the one or more RNA species are identified by amplifying cDNA with a plurality of primers comprising a primer in a 5′ Immunoglobulin E (IgE) constant region exon and either a primer in the Is-exon or a primer in the epsilon switch region (Se) to thereby detect GLT RNA species comprising one or more nucleic acid sequences containing a splice junction spanning the 5′ IgE constant region exon and the Is-exon or the Se; and

predicting the future allergic response to the one or more allergens in the subject when an amount of the measured and identified &GLT RNA species exceeds a threshold, wherein the threshold is one determined by measuring the amounts of measured and identified ¿GLT RNA species in patients with a known allergic condition.

2 . The method of claim 1 , wherein the predicted future allergic response is a predicted predisposition to developing an allergy to the one or more allergens.

3 . The method of claim 1 , wherein the predicted future allergic response is a prediction of allergic reaction severity to the one or more allergens.

4 . The method of claim 1 , further comprising the step of predicting alleviation or elimination of an existing allergic response to the one or more allergens.

5 . The method of claim 4 , further comprising the step of determining an appropriate therapeutic intervention for the allergy.

6 . The method of claim 1 , wherein the predicting step comprises identifying at least one allergen from the one or more allergens that causes the predicted future allergic response.

7 . The method of claim 6 , wherein the allergen comprises at least one of a food allergen, a plant allergen, a fungal allergen, an animal allergen, a drug allergen, a cosmetic allergen, and a latex allergen.

8 . The method of claim 1 , wherein the one or more RNA species is one produced by germline transcription.

9 . The method of claim 1 , wherein the one or more nucleic acid sequences contain a splice junction comprising the 5′ IgE constant region exon and the Iε-exon.

10 . The method of claim 1 , wherein the one or more nucleic acid sequences contain a splice junction comprising the 5′ IgE constant region exon is and the Sε.

11 . The method of claim 1 , wherein the measuring and identifying step further comprises sequencing the one or more RNA species.

12 . The method of claim 11 , wherein the sequencing step comprises bulk RNA-Seq, single-cell RNA-Seq, direct RNA sequencing, or in-situ hybridization.

13 . The method of claim 1 , wherein amplifying the cDNA further comprises using biotinylated capture probes.

14 . The method of claim 1 , wherein the measuring and identifying step further comprises reverse transcription-quantitative PCR.

15 . The method of claim 1 , wherein enriching the B cells comprises fluorescence activated cell sorting (FACS).

16 . The method of claim 1 , wherein enriching the B cells comprises depleting the blood sample of non-B cells.

17 . The method of claim 1 , wherein enriching the B cells comprises centrifuging the blood sample, resuspending the pelleted cells, and subjecting the resuspended cells to density centrifugation.

Assignments (1)

Continuity (2)

References Cited (125)