- Document Number:
20240368713
- Appl. No:
18/777038
- Application Filed:
July 18, 2024
- نبذة مختصرة :
The present invention relates to the use of the measure of anelloviral load for the determination of immunosuppression. More precisely, the present invention provides a method for characterizing the immunosuppressed or non-immunosuppressed status of a subject, comprising the steps of determining the anelloviral load from a biological sample of the said subject, and determining from the said comparison the immunosuppressed or non-immunosuppressed status. The determination of the immunosuppressed status of the subject can then be used to design or adapt a therapeutic treatment.
- Assignees:
INSTITUT PASTEUR (Paris, FR), UNIVERSITE PARIS CITE (PARIS, FR), ASSISTANCE PUBLIQUE - HOPITAUX DE PARIS (Paris, FR), PATHOQUEST (Paris, FR), ECOLE NATIONALE VETERINAIRE D' ALFORT (Maison Alfort, FR)
- Claim:
1. A method, comprising: 1) obtaining a biological sample from a human subject who is or is suspected of being immunosuppressed; and 2) measuring the number of anellovirus DNA sequences in the sample by a method comprising detecting Torque teno virus (TTV) DNA, Torque teno midi virus (TTMDV) DNA, and Torque teno mini virus (TTMV) DNA, wherein measuring the number of anellovirus DNA sequences in the sample comprises at least one amplification step or sequencing step with a primer comprising at least 12 consecutive bases of SEQ ID NO: 4, which hybridizes specifically to TTV, TTMDV, and TTMV DNA within position +121 to +151 relative to the TATA box consensus or at least 12 consecutive bases of SEQ ID NO: 5, which hybridizes specifically to TTV, TTMDV, and TTMV DNA within position +20 to +53 relative to the TATA box consensus, with the proviso that the primer does not have the sequence of SEQ ID No. 6 or SEQ ID No. 7, using a nucleic acid amplification instrument and/or a sequencing instrument; 3) measuring the number of total DNA sequences in the sample, wherein measuring the number of total DNA sequences in the sample comprises using a nucleic acid amplification instrument and/or a sequencing instrument; and 4) generating a ratio of the measured number of anellovirus DNA sequences in the sample to the measured number of total DNA sequences in the sample, wherein the ratio of the measured number of anellovirus DNA sequences in the sample to the measured number of total DNA sequences in the sample is higher than 0.2×10−4 in the sample; responsive to measuring the ratio at higher than 0.2×10−4, determining that the human subject is immunosuppressed.
- Claim:
2. The method of claim 1, wherein said human subject suffers from transplant rejection.
- Claim:
3. The method of claim 1, wherein said human subject suffers from an infection.
- Claim:
4. The method of claim 1, wherein the number of anellovirus DNA sequences is measured by hybridization with a labeled probe, PCR amplification or sequencing.
- Claim:
5. The method of claim 1, wherein the number of anellovirus DNA sequences is measured by sequencing said anellovirus DNA.
- Claim:
6. The method of claim 1, wherein the number of anellovirus DNA sequences is measured by a method comprising at least one amplification step with primers comprising at least 15 consecutive bases of SEQ ID NO: 4 or 5.
- Claim:
7. The method of claim 6 wherein said primers further comprise at least one of: a functional group for covalent coupling at the 5′ or 3′ end, a spacer molecule or sequence at the 5′ or 3′ end, additional sequences as index or tag sequences to perform pre or post additional and general amplification steps not depending on the target sequences to be quantified.
- Claim:
8. The method of claim 1, wherein the number of anellovirus DNA sequences is measured by massive parallel sequencing.
- Claim:
9. The method of claim 1, wherein measuring the number of anellovirus DNA sequences in the sample further comprises the steps of assigning each anellovirus sequence to a specific anellovirus genome and numbering the copies of anellovirus genomes thus identified.
- Claim:
10. The method of claim 1, wherein the number of anellovirus DNA sequences in the sample is determined by quantitative PCR.
- Claim:
11. The method of claim 1, wherein measuring the number of anellovirus DNA sequences in the sample comprises the amplification of at least one consensus anellovirus sequence.
- Claim:
12. The method of claim 7 wherein said functional group for covalent coupling at the 5′ or 3′ end is a terminal group comprising a thiol, amine or carboxyl group.
- Claim:
13. The method of claim 1, wherein determining that the human subject is immunosuppressed further comprises determining a degree of immunosuppression for the subject.
- Claim:
14. A method of preparing a value indicative of an immunosuppressed status in a human subject who is or is suspected of being immunosuppressed, comprising: 1) obtaining a biological sample from the human subject; 2) using a nucleic acid amplification instrument and/or a sequencing instrument with a primer comprising at least 15 consecutive bases of SEQ ID NO: 4 or SEQ ID NO: 5, with the proviso that the primer does not have the sequence of SEQ ID No. 6 or SEQ ID No. 7 to measure the number of anellovirus DNA sequences in the sample by detecting Torque teno virus (TTV) DNA, Torque teno midi virus (TTMDV) DNA, and Torque teno mini virus (TTMV) DNA in the sample to generate a combined value of the measured number of anellovirus; and 3) using a nucleic acid amplification instrument and/or a sequencing instrument to measure the number of total DNA sequences in the sample; 4) generating the value by determining a ratio of the measured number of anellovirus DNA sequences to the measured number of total DNA sequences in the sample, wherein if the value is higher than 0.2×10−4 in the sample, the human subject is immunosuppressed.
- Claim:
15. The method of claim 14, wherein PCR is used to obtain the measured number of anellovirus DNA sequences using a pair of primers comprising at least 15 consecutive bases of SEQ ID NO: 4 or 5.
- Claim:
16. The method of claim 15, further comprising a labelled probe that hybridizes between the pair of primers.
- Claim:
17. The method of claim 1, wherein the sample is a blood sample.
- Claim:
18. The method of claim 14, wherein the sample is a blood sample.
- Current International Class:
12
- الرقم المعرف:
edspap.20240368713
Processing Request
No Comments.