US 12,467,094 B2
Methods and systems for predicting allergic response
Derek Croote, South San Francisco, CA (US)
Assigned to IGGENIX, INC., South San Francisco, CA (US)
Filed by IgGenix, Inc., South San Francisco, CA (US)
Filed on Mar. 11, 2022, as Appl. No. 17/692,408.
Claims priority of provisional application 63/159,889, filed on Mar. 11, 2021.
Prior Publication US 2022/0290240 A1, Sep. 15, 2022
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6851 (2018.01); C12Q 1/6883 (2018.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/6851 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/158 (2013.01)] 17 Claims
 
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.