# US 11,299,788 B2 · Method for the quantification of Faecalibacterium prausnitzii phylogroup I and/or phylogroup II members and the use thereof as biomarkers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,299,788 B2 - Document ID: US11299788B2 - Status: Granted - Kind: B2 - Application number: 15/751,805 - Filed: 2016-08-11 - Published: 2019-01-24 - Granted: 2022-04-12 - Earliest claimed: 2015-08-11 - Assignees: Universitat De Girona; Fundacio Institut D'Investigacio Biomedica de Girona Dr. Josep Trueta; Goodgut S.L. - Applicants: Universitat de Girona; Fundado Institut D'Investigacio Biomedica de Girona Dr. Josep Trueta; Goodgut S.L. - Inventors: Mireia Lopez Siles; Librado Jesus Garcia Gil; Xavier Aldeguer Mante; Margarita Martinez Medina - Web page: https://patentgrep.com/patent/US11299788B2 ## CPC - C12Q 1/689: for bacteria - C12Q 1/6883: for diseases caused by alterations of genetic material - C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575) - C12Q 2600/106: Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism - C12Q 2600/112: Disease subtyping, staging or classification - C12Q 2600/118: Prognosis of disease development - C12Q 2600/158: Expression markers - Y02A 90/10: Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation - 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 - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract The present invention relates to a novel method for an accurate quantification in intestinal samples of Faecalibacterium prausnitzii phylogroup I members (PHGI) and/or Faecalibacterium prausnitzii phylogroup II members (PHGII). It further relates to a method for detecting intestinal diseases, including the screening, diagnosis, differential diagnosis, and/or monitoring of disease activity or progression in a human subject comprising determining the abundance of PHGI and/or PHGII in an intestinal sample from said subject. Moreover, it relates to a method for the prediction of the efficacy of a drug in the therapeutic treatment of an intestinal disease in a human subject comprising determining the abundance of PHGI and/or PHGII in an intestinal sample from said subject. ## Claims ### Claim 1 An in vitro method for determining the abundance of one or more of (a) Faecalibacterium prausnitzii phylogroup I members (PHGI) and (b) Faecalibacterium prausnitzii phylogroup II members (PHGII) in an intestinal sample from a subject by quantitative Polymerase Chain Reaction (qPCR) comprising F. prausnitzii 16S rRNA gene amplification comprising using: (i) F. prausnitzii species-specific 16S rRNA gene primers; and (ii) (a) a PHGI-specific probe consisting of the nucleotide sequence of SEQ ID NO:3 for determining the abundance of PHGI; and/or (b) a PHGII-specific probe consisting of the nucleotide sequence of SEQ ID NO:4 for determining the abundance of PHGII. ### Claim 2 (depends on 1) The method of claim 1, wherein the intestinal sample from the subject comprises a fecal sample, and wherein the method further comprises comparing: (a) a value selected from: (i) PHGII abundance; (ii) PHGI and/or PHGII abundance; and/or (iii) a ratio of PHGII abundance to E. coli (EC) abundance in the subject sample; with (b) a corresponding values in a reference sample, wherein a significant deviation in the subject sample value with regard to the corresponding value in the reference sample is indicative of inflammatory bowel disease (IBD). ### Claim 3 (depends on 2) The method according to claim 2, wherein the reference sample is a healthy subject sample. ### Claim 4 (depends on 2) The method according to claim 2, wherein the reference sample is a previous sample of the same subject. ### Claim 5 (depends on 1) The method according to claim 1, wherein PHGI abundance determination is performed with a forward primer with at least 75% identity to SEQ ID NO: 1, a reverse primer with at least 75% identity to SEQ ID NO: 2, and the PHGI-specific probe consisting of the nucleotide sequence of SEQ ID NO:3. ### Claim 6 (depends on 1) The method according to claim 1, wherein PHGII abundance determination is performed with: a forward primer comprising the nucleotide sequence of SEQ ID NO: 1, a reverse primer comprising the nucleotide sequence of SEQ ID NO: 2; and the PHGII-specific probe consisting of the nucleotide sequence of SEQ ID NO: 4. ### Claim 7 (depends on 1) The method according to claim 1, wherein the PHGI and/or PHGII abundance levels are normalized, wherein normalization is carried out (i) with respect to total bacteria quantification or (ii) by DNA concentration. ### Claim 8 (depends on 1) The method according to claim 1, wherein the intestinal sample from the subject is a feces sample. ### Claim 9 (depends on 1) The method according to claim 1, wherein the method further comprises detecting and/or quantifying one or more additional biomarkers of intestinal disease and optionally comprises combining the results of PHGI abundance, PHGII abundance and/or additional biomarkers of intestinal disease. ### Claim 10 (depends on 1) The method of claim 1, further comprising determining the abundance of PHGI and PHGII in the intestinal sample from the subject. ### Claim 11 (depends on 1) The method of claim 1, wherein the intestinal sample from the subject is a fecal sample, and wherein the method further comprises comparing: (a) the PHGII abundance in the subject sample; with (b) a corresponding value in a reference sample, wherein a significant deviation in the PHGII abundance in the subject sample value with regard to the corresponding value in the reference sample is indicative of inflammatory bowel disease (IBD). ### Claim 12 (depends on 1) The method of claim 1, wherein the intestinal sample from the subject is a fecal sample, the method further comprises comparing: (a) a value selected from: (1) the PHGII abundance, or (2) the PHGI and PHGII abundance in the subject sample; with (b) a corresponding value in a reference sample, wherein a significant deviation in the subject sample value with regard to the corresponding value in the reference sample is indicative of inflammatory bowel disease (IBD). ### Claim 13 (depends on 1) The method of claim 1, wherein the intestinal sample from the subject comprises a fecal sample, and wherein the method further comprises comparing: (a) a value selected from: (1) the PHGII abundance, (2) a ratio of the PHGII abundance to E. coli (EC) abundance, in the subject sample; with (b) a corresponding value in a reference sample, wherein a significant deviation in the subject sample value with regard to the corresponding value in the reference sample is indicative of inflammatory bowel disease (IBD). ### Claim 14 (depends on 1) The method of claim 1, wherein the intestinal sample from the subject comprises a fecal sample, and wherein the method further comprises comparing: (a) the PHGII abundance in the subject sample with (b) a corresponding value in a reference sample, wherein a decrease in the PHGII abundance in the subject sample with regard to the corresponding value in the reference sample is indicative of IBD. ### Claim 15 (depends on 1) The method of claim 1, wherein the PHGII abundance is determined by quantitative PCR on the intestinal sample from the subject, wherein the intestinal sample is a feces sample, and is expressed as a Ct value, and wherein the method further comprises comparing: (a) the PHGII Ct value with (b) a cut-off Ct value, wherein an increase in the PHGII Ct value in the feces sample of the subject with respect to the cut-off Ct value is indicative of IBD.