Method for diagnosis and monitoring of viral infection by analysis of viral transrenal nucleic acids in urine
The present invention relates to methods for diagnosis or monitoring of viral infection by detecting the presence of transrenal viral nucleic acids or nucleic acids of viral origin in urine sample, with or without isolation of nucleic acids from a urine sample. The analysis of the nucleic acids is performed through hybridization of the nucleic acids with specific probes, or through a chain amplification reaction with specific primers. The methods are applicable to all viral pathogenic agents, including RNA, DNA, episomal, or integrated viruses.
1. A method for diagnosing a virus infection in a subject, comprising
a) obtaining a urine sample from a subject;
b) separating a soluble fraction of said urine sample; and
c) detecting the presence of a cell-free virus nucleic acid in said soluble fraction of said urine sample, wherein said nucleic acid is less than about 300 bp in length, wherein said detecting is performed by a method selected from the group consisting of PCR, nested PCR, SSCP, LCR, and SDA, using a primer set comprising
at least one forward primer selected from the group consisting of SEQ ID NOs: 1 and 3 and at least one reverse primer selected from the group consisting of SEQ ID NOs: 2 and 4, or
a forward primer of SEQ ID NO: 5 and at least one reverse primer selected from the group consisting of SEQ ID NOs: 6 and 7, or
a forward primer of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 10,
wherein the presence of said nucleic acid diagnoses a viral infection.
2. The method of claim 1 , further comprising the step of quantifying the nucleic acid.
3. The method of claim 1 , wherein the subject is a mammal.
4. The method of claim 3 , wherein the mammal is a human.
5. A method for diagnosing a virus infection in a subject, comprising
a) obtaining a urine sample from a subject;
b) separating a soluble fraction of said urine sample; and
c) detecting the presence of a cell-free virus nucleic acid in said soluble fraction of said urine sample by a method selected from the group consisting of PCR, nested PCR, SSCP, LCR, and SDA, using a forward primer of SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 8, wherein the presence of said nucleic acid diagnoses a viral infection.
6. The method of claim 5 , further comprising the step of quantifying the nucleic acid.
7. The method of claim 5 , wherein the subject is a mammal.
8. The method of claim 7 , wherein the mammal is a human.
9. A method for diagnosing a virus infection in a subject, comprising
a) obtaining a urine sample from a subject;
b) separating a soluble fraction of said urine sample; and
c) detecting the presence of a cell-free virus nucleic acid in said soluble fraction of said urine sample by a method selected from the group consisting of PCR, nested PCR, SSCP, LCR, and SDA, using a forward primer of SEQ ID NO: 9 and a reverse primer of SEQ ID NO: 10, wherein the presence of said nucleic acid diagnoses a viral infection.
10. The method of claim 9 , further comprising the step of quantifying the nucleic acid.
11. The method of claim 9 , wherein the subject is a mammal.
12. The method of claim 11 , wherein the mammal is a human.
13. The method of claim 2 , wherein said primer set comprises at least one forward primer selected from SEQ ID NOs: 1 and 3 and at least one reverse primer selected from SEQ ID NOs: 2 and 4.
14. The method of claim 4 , wherein said primer set comprises at least one forward primer selected from SEQ ID NOs: 1 and 3 and at least one reverse primer selected from SEQ ID NOs: 2 and 4.
15. The method of claim 2 , wherein said primer set comprises a forward primer of SEQ ID NO: 5 and at least one reverse primer selected from the group consisting of SEQ ID NOs: 6 and 7.
16. The method of claim 4 , wherein said primer set comprises a forward primer of SEQ ID NO: 5 and at least one reverse primer selected from the group consisting of SEQ ID NOs: 6 and 7.
17. The method of claim 2 , wherein said primer set comprises a forward primer of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 10.
18. The method of claim 4 , wherein said primer set comprises a forward primer of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 10.