# US 10,370,719 B2 · Method and device for diagnosing organ injury > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,370,719 B2 - Document ID: US10370719B2 - Status: Granted - Kind: B2 - Application number: 15/526,296 - Filed: 2015-11-12 - Published: 2017-11-23 - Granted: 2019-08-06 - Earliest claimed: 2014-11-12 - Assignees: Hitachi Chemical Company, Ltd.; Hitachi Chemical Company America, Ltd.; The Regents of the University of California - Applicants: Hitachi Chemical Co., Ltd.; Hitachi Chemical Co. America, Ltd.; The Regents of the University of California - Inventors: Satish P. RamachandraRao; Ravindra Lall Mehta; Masato Mitsuhashi; Taku Murakami - Web page: https://patentgrep.com/patent/US10370719B2 ## CPC - C12Q 1/6883: for diseases caused by alterations of genetic material - C12Q 2600/118: Prognosis of disease development - C12Q 2600/158: Expression markers - 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 ## Abstract The present disclosure relates to methods of collecting exosomes and microvesicles (EMV) from urine and isolating corresponding mRNA in order to diagnose and treat acute kidney injury (AKI). In particular, certain embodiments relate to the method of capturing EMV from urine applied to a filter device that is capable of capturing EMV. Nucleic acids such as mRNA can be isolated from the EMV using an oligo(dT)-coated plate designed to accommodate the filter device and then used for further molecular analysis. Quantification of the collected nucleic acids may then be used in the diagnosis and/or treatment of IBD. ## Claims ### Claim 1 A method of treating an acute kidney injury (AKI) in a human subject, the method comprising: (A) having a vesicle-containing sample obtained from said human subject sent to a laboratory for the laboratory to perform an assay comprising the following steps (1)-(3): (1) capturing at least a portion of vesicles from said sample on or in a vesicle-capture material, thereby generating a vesicle sample; (2) quantifying an expression level of at least one gene in said vesicle sample, wherein said at least one gene comprises CALM1 and at least one additional gene selected from the group consisting of SLC12A1, CFLAR, GSTA1, HIF1A, and ALB; (3) determining that said subject has said AKI due to said expression level of each of said at least one gene being significantly different from the expression level of the respective gene in a vesicle sample of a healthy human control subject not suffering from said AKI, thereby diagnosing said subject as having AKI; and (B) administering an effective amount of an AKI medication to the human subject having said AKI, wherein said AKI medication is selected from the group consisting of a diuretic agent, an intravenous fluid, a steroid medication, a plasma exchange, and a cyclophosphamide, thereby treating said AKI in said human subject. ### Claim 2 (depends on 1) The method of claim 1, wherein said vesicle-containing sample is a urine sample. ### Claim 3 (depends on 1) The method of claim 1, wherein capturing comprises passing said vesicle-containing sample through a filter comprising glass fiber. ### Claim 4 (depends on 1) The method of claim 1, further comprising lysing said vesicle sample on or in said vesicle-capture material. ### Claim 5 (depends on 1) The method of claim 1, wherein quantifying an expression level of at least one gene in said vesicle sample comprises quantifying said expression level of three genes. ### Claim 6 (depends on 1) The method of claim 1, wherein capturing further comprises: receiving said fluid sample; and passing said fluid sample through a vesicle capture material adapted to capture a captured vesicle population within said vesicle capture material. ### Claim 7 (depends on 6) The method of claim 6, wherein quantifying comprises: positioning said vesicle capture material and said captured vesicle population adjacent to a substrate comprising immobilized oligo(dT); applying lysis buffer to said vesicle capture material, thereby lysing said captured vesicle population; hybridizing one or more mRNA from said captured vesicle population to said substrate; synthesizing directly on said substrate one or more cDNA from said one or more mRNA; and quantifying by PCR analysis of said one or more cDNA an expression level of a marker mRNA in said captured vesicle population. ### Claim 8 (depends on 6) The method of claim 6, wherein receiving comprises receiving said fluid sample from a medical professional. ### Claim 9 (depends on 7) The method of claim 7, wherein said vesicle capture material comprises glass fiber. ### Claim 10 (depends on 9) The method of claim 9, wherein passing comprises loading said fluid sample into a device comprising a loading reservoir removably attached to a tip, wherein said vesicle capture material is housed within said tip. ### Claim 11 (depends on 1) The method of claim 1, wherein determining further comprises: determining a gene classifier by summing the expression level of at least two of the at least one genes; and determining whether the gene classifier is significantly different from a respective gene classifier in a vesicle sample of a healthy human control subject not suffering from said AKI. ### Claim 12 (depends on 11) The method of claim 11, wherein the gene classifier comprises the expression levels of at least CALB1 and CALM1. ### Claim 13 (depends on 12) The method of claim 12, wherein the gene classifier further comprises the expression level of GAPDH.