# US 12,215,386 B2 · Blood biomarkers for appendicitis and diagnostics methods using biomarkers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,215,386 B2 - Document ID: US12215386B2 - Status: Granted - Kind: B2 - Application number: 17/343,285 - Filed: 2021-06-09 - Published: 2022-03-24 - Granted: 2025-02-04 - Earliest claimed: 2014-10-22 - Assignees: The George Washington University - Applicants: The George Washington University - Inventors: Lakhmir S. Chawla; Timothy A. McCaffrey - Web page: https://patentgrep.com/patent/US12215386B2 ## CPC - C12Q 1/6883: for diseases caused by alterations of genetic material - G01N 33/6893: {related to diseases not provided for elsewhere} - C12Q 2600/158: Expression markers - G01N 33/566: using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} - G01N 33/84: involving inorganic compounds or pH - G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A) - G01N 2800/06: Gastro-intestinal diseases - G01N 2800/56: Staging of a disease; Further complications associated with the disease - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The invention relates to methods and kits for diagnosing and/or treating appendicitis in a subject, comprising obtaining a biological sample rom said subject; detecting RNA expression levels of at least three or more biomarkers in the biological sample and comparing the expression levels of said at least three or more biomarkers to a control sample wherein an increase or decrease in the level of expression of said at least three or more biomarkers as compared to the control sample is indicative of appendicitis. ## Claims ### Claim 1 A method of treating a human subject diagnosed with appendicitis comprising administering an antibiotic to the subject, removing the appendix of the subject, or a combination thereof, wherein the human subject is diagnosed by a method comprising: obtaining a biological sample from said subject; detecting expression levels of at least three or more biomarkers in the biological sample selected from the group consisting of Chemokine C-X-C receptor 1, Interleukin 8 receptor β, Fe frag of IgG receptor IIIb (CD16b), MHC class II DR beta 5, Leukocyte IgG-like receptor A3, Defensin alpha 1, Defensin alpha 1B, Defensin alpha 3, 18S ribosomal RNA, CDC14A, 28S ribosomal RNA, 60S acidic ribosomal protein P1, 40S ribosomal protein S26, Ribosomal protein L23, Ribosomal protein L37a, Ribosomal protein S28, Alkaline phosphatase, Carbonic anhydrase IV, Neuroblastoma breakpoint family 10, Ninjurin 1, Prokineticin 2, Superoxide dismutase 2, LOC100129902, LOC100131205, LOC100131905, LOC100132291, LOC100132394, LOC100132742, LOC100134364, LOC391370, LOC646785, LOC644191 and C5orf32; and diagnosing the subject with appendicitis when at least three or more of said biomarkers are differentially expressed compared to the expression levels of said biomarkers in a group of human subjects not suffering from appendicitis. ### Claim 2 (depends on 1) The method of claim 1, wherein said biological sample is a blood sample, serum sample, or a plasma sample. ### Claim 3 (depends on 1) The method of claim 1, wherein said at least three biomarkers are selected from the list consisting of Interleukin 8 receptor β, Defensin alpha 1, Defensin alpha 1B, Defensin alpha 3 and Alkaline phosphatase. ### Claim 4 (depends on 1) The method of claim 1, further comprising determining the ratio of expression between Alkaline phosphatase or Interleukin 8 receptor β and Defensin alpha 1, Defensin alpha 1B or Defensin alpha 3 and comparing said ratio to a control sample, wherein an increase in the ratio of expression is indicative of appendicitis. ### Claim 5 (depends on 1) The method of claim 1, wherein detecting expression levels of at least three or more biomarkers comprises measuring RNA levels of said at least three or more biomarkers. ### Claim 6 (depends on 5) The method of claim 5, wherein measuring RNA levels comprises using reverse transcriptase and polymerase chain reaction. ### Claim 7 (depends on 5) The method of claim 5, wherein measuring RNA levels comprises using microarray analysis. ### Claim 8 (depends on 5) The method of claim 5, wherein measuring RNA levels comprises using RNA sequencing. ### Claim 9 (depends on 1) The method of claim 1, wherein comparing the expression levels of said at least three or more biomarkers comprises using a prediction model to determine if a pattern of expression of said at least three or more biomarkers is indicative of appendicitis. ### Claim 10 (depends on 5) The method of claim 5, wherein said at least three RNA sequences are selected from the group consisting of LOC100129902, LOC100131205, LOC100131905, LOC100132291, LOC100132394, LOC100132742, LOC100134364, LOC391370, LOC646785, LOC644191 and C5orf32. ### Claim 11 (depends on 1) The method of claim 1, further comprising performing a computed tomography (CT) scan, a magnetic resonance imaging (MRI) scan, an ultrasound sonography, or any combination thereof on said subject prior to treating said subject for appendicitis. ### Claim 12 (depends on 5) The method of claim 5, wherein said three RNA sequences are Interleukin 8 receptor β, Defensin alpha 1, and Alkaline phosphatase. ### Claim 13 A method of treating a human subject diagnosed with appendicitis comprising administering an antibiotic to the subject, removing the appendix of the subject, or a combination thereof, wherein the human subject is diagnosed by a method comprising: obtaining a biological sample from said subject; detecting the expression level of Defensin alpha 3; and diagnosing the subject with appendicitis when the Defensin alpha 3 is differentially expressed compared to the expression level of Defensin alpha 3 in a group of human subjects not suffering from appendicitis. ### Claim 14 (depends on 13) The method of claim 13, wherein detecting the expression level comprises measuring RNA level of Defensin alpha 3. ### Claim 15 (depends on 14) The method of claim 14, wherein measuring RNA levels comprises using reverse transcriptase and polymerase chain reaction. ### Claim 16 (depends on 14) The method of claim 14, wherein measuring RNA levels comprises using microarray analysis. ### Claim 17 (depends on 14) The method of claim 14, wherein measuring RNA levels comprises using RNA sequencing.