Claims for Patent: 9,631,243

Summary for Patent: 9,631,243

Title:	Methods and compositions for identifying JC virus
Abstract:	The disclosure relates to methods and compositions for detecting the presence of Prototype and/or Archetype JC virus in a biological sample from a subject. In some embodiments, the methods include amplifying and detecting a first nucleic acid sequence unique to Archetype JC virus in a biological sample, and a second nucleic acid sequence common to both Archetype and Prototype JC virus in the biological sample. In several embodiments, the methods can be used to identify JC virus in a biological sample from a subject at risk for progressive multifocal leukoencephalopathy. Compositions and kits for use in the disclosed methods are also provided.
Inventor(s):	Major; Eugene (Darnestown, MD), Ryschkewitsch; Caroline (Chevy Chase, MD)
Assignee:	THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES (Washington, DC)
Application Number:	14/408,919
Patent Claims:	1. A method of detecting the presence or absence of Prototype JC virus in a biological sample from a subject, comprising: amplifying and detecting an amount of a first nucleic acid molecule and an amount of a second nucleic acid molecule from the biological sample using a using a multiplex quantitative real-time polymerase chain reaction (qPCR) assay, comprising: amplifying the first nucleic acid molecule with a first oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of the nucleotide sequences set forth as SEQ ID NOs: 3 (JRR-1) and 4 (JRR-2), respectively; detecting the amplified first nucleic acid molecule with a first oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 5 (JRR-1.1), or the complement thereof; amplifying the second nucleic acid molecule with a second oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of nucleotide sequences set forth as SEQ ID NOs: 6 (JCT-3) and 7 (JCT-4), respectively; detecting the amplified second nucleic acid molecule with a second oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 8 (JCT-1.2), or the complement thereof; and wherein the first nucleic acid molecule is present in an Archetype JC virus genome but not the Prototype JC virus genome, and the second nucleic acid molecule is present in both the Archetype and Prototype JC virus genomes; detecting a greater amount of the second nucleic acid molecule than the first nucleic acid molecule in the biological sample identifies the presence of Prototype JC virus in the biological sample; and detecting an equal amount of the first and second nucleic acid molecules in the biological sample identifies the absence of Prototype JC virus in the biological sample. 2. The method of claim 1, wherein the forward and reverse oligonucleotide primers of the first oligonucleotide pair consist of the nucleotide sequences set forth as SEQ ID NOs: 3 and 4, respectively; and the forward and reverse oligonucleotide primers of the second oligonucleotide pair consist of the nucleotide sequences set forth as SEQ ID NOs: 6 and 7, respectively. 3. The method of claim 1, wherein the subject is a subject at risk of developing progressive multifocal leukoencephalopathy (PML). 4. The method of claim 3, wherein subject is an immunocompromised subject. 5. The method of claim 3, wherein the subject has an HIV infection or multiple sclerosis. 6. The method of claim 5, wherein the subject has AIDS. 7. The method of claim 3, wherein the subject is being treated with an immunosuppressive therapy. 8. The method of claim 7, wherein the subject is being treated with natalizumab. 9. The method of claim 1, wherein the biological sample is a cerebrospinal fluid (CSF), blood or urine sample. 10. The method of claim 1, wherein detecting the presence of Prototype JC virus in the biological sample identifies the subject as having an increased risk of developing progressive multifocal leukoencephalopathy (PML). 11. The method of claim 10, further comprising inhibiting PML in the subject, comprising administering to the subject a therapeutically effective amount of an anti-PML agent, thereby inhibiting PML in the subject. 12. The method of claim 11, wherein the anti-PML agent is cytosine arabinoside or cidofovir. 13. A composition, comprising: a first oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of the nucleotide sequences set forth as SEQ ID NOs: 3 and 4, respectively; and a first oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 5, or the complement thereof, wherein the first oligonucleotide probe is labeled with a detectable marker. 14. The composition of claim 13, wherein the forward and reverse oligonucleotide primers of the first oligonucleotide primer pair consist of the nucleotide sequences set forth as SEQ ID NOs: 3 and 4, respectively. 15. The composition of claim 13, further comprising: a second oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of the nucleotide sequences set forth as SEQ ID NOs: 6 and 7, respectively; and a second oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 8, or the complement thereof; wherein the second oligonucleotide probe is labeled with a detectable marker that is different from the detectable marker of the first oligonucleotide probe. 16. The composition of claim 15, wherein the forward and reverse oligonucleotide primers of the first oligonucleotide primer pair consist of the nucleotide sequences set forth as SEQ ID NOs: 3 and 4, respectively; and the forward and reverse oligonucleotide primers of the second oligonucleotide primer pair consist of the nucleotide sequences set forth as SEQ ID NOs: 6 and 7, respectively. 17. A kit for detecting the presence of Prototype and/or Archetype JC virus in a biological sample from a subject, comprising: a container comprising the composition of claim 13; and instructions for using the kit. 18. A kit for detecting the presence of Prototype and/or Archetype JC virus in a biological sample from a subject, the kit comprising a set of containers comprising: a first oligonucleotide primer consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 3; a second oligonucleotide primer consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 4; a third oligonucleotide primer consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 6; a fourth oligonucleotide primer consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 7; a first oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 5, or the complement thereof; a second oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 8, or the complement thereof; and wherein the first and second oligonucleotide probes are labeled with different detectable labels. 19. The kit of claim 18, wherein the first, second, third, and fourth oligonucleotide primers consist of the nucleotide sequences set forth as SEQ ID NOs: 3, 4, 6, and 7, respectively.

Details for Patent 9,631,243

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Biogen Inc.	TYSABRI	natalizumab	Injection	125104	11/23/2004		2032-06-18