Claims for Patent: 8,859,748

Summary for Patent: 8,859,748

Title:	Nucleic acid amplification primers for PCR-based clonality studies
Abstract:	The invention relates to PCR-based clonality studies for among others early diagnosis of lymphoproliferative disorders. Provided is a set of nucleic acid amplification primers comprising a forward primer, or a variant thereof, and a reverse primer, or a variant thereof, capable of amplifying a rearrangement selected from the group consisting of a VH-JH IGH rearrangement, a DH-JH IGH rearrangement, a VK-JK IGK rearrangement, a VK/intron-Kde IGK rearrangement, a V.lamda.-J.lamda. IGL rearrangement, a V.beta.-J.beta. TCRB rearrangement, a D.beta.-J.beta. TCRB rearrangement, a V.gamma.-J.gamma. TCRG rearrangement, a V.delta.-J.delta. TCRD rearrangement, a D.delta.-D.delta. TCRD rearrangement, a D.delta.-J.delta. TCRD rearrangement, a V.delta.-D.delta. TCRD rearrangement, or a translocation selected from t(11;14)(BCL1-IGH) and t(14;18)(BCL2-IGH). The primers can be used in PCR-based clonality studies for early diagnosis of lymphoproliferative disorders and detection of minimal residual disease (MRD). Also provided is a kit comprising at least one set of primers of the invention.
Inventor(s):	Van Dongen; Jacobus Johannes Maria (Vorden, NL), Langerak; Anthonie Willem (Barendrecht, NL), Schuuring; Eduardus Maria Dominicus (Groningen, NL), San Miquel; Jesus Fernando (Salamanca, ES), Garcia Sanz; Ramon (Salamanca, ES), Parreira; Antonio (Lisbon, PT), Smith; John Lewis (Wilts, GB), Lavender; Frances Louise (Hants, GB), Morgan; Gareth John (Surrey, GB), Evans; Paul Anthony Stuart (West Yorkshire, GB), Kneba; Michael (Westensee, DE), Hummel; Michael (Berlin, DE), Macintyre; Elizabeth Anne (Meudon, FR), Bastard; Christian (Ardouval, FR)
Assignee:
Application Number:	10/531,106
Patent Claims:	1. A kit for the detection of at least one IGH rearrangement, wherein said kit comprises a plurality of forward primers selected from the group consisting of SEQ ID NOs: 1-8, 10-15, 17-20, and 22-28, and at least one reverse primer comprising SEQ ID NO:21, wherein at least one of said plurality of forward primers or at least one of said at least one reverse primer comprises a fluorescent label. 2. The kit of claim 1 further comprising forward primers comprising SEQ ID NOs: 9 and 16. 3. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.sub.K-J.sub.K IGK rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 29-34 and one or more reverse primers of a sequence selected from SEQ ID NOs: 35 and 36. 4. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.sub.K/intron-Kde IGK rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 29-34, 38 and a reverse primer of SEQ ID NO: 37. 5. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.lamda.-J.lamda. IGL rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 39 and 40 and a reverse primer of SEQ ID NO: 41. 6. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.beta.-J.beta. TCRB rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 42-64 and one or more reverse primers of a sequence selected from SEQ ID NOs: 65-77. 7. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a D.beta.-J.beta. TCRB rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 78 and 79 and one or more reverse primers of a sequence selected from SEQ ID NOs: 65-77. 8. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.sub..gamma.-J.sub..gamma. TCRG rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 80-83 and one or more reverse primers of a sequence selected from SEQ ID NOs: 84 and 85. 9. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.delta.-J.delta. TCRD rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 86-91 and one or more reverse primers of a sequence selected from SEQ ID NOs: 92-95. 10. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a D.delta.-D.delta. TCRD rearrangement comprising a forward primer of SEQ ID NO: 96 and a reverse primer of SEQ ID NO: 97. 11. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a D.delta.-J.delta. TCRD rearrangement comprising a forward primer of SEQ ID NO: 96 and one or more reverse primers of a sequence selected from SEQ ID NOs: 92-95. 12. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a V.delta.-D.delta. TCRD rearrangement comprising one or more forward primers of a sequence selected from SEQ ID NOs: 86-91 and a reverse primer of SEQ ID NO: 97. 13. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a chromosomal translocation (11;14) (BCL1-IGH) comprising a forward primer of SEQ ID NO: 98 and a reverse primer of SEQ ID NO: 21. 14. The kit of claim 1, further comprising a set of nucleic amplification primers capable of amplifying a chromosomal translocation t(14;18) (BCL2-IGH), comprising one or more forward primers of a sequence selected from SEQ ID NO: 100-109 and a reverse primer of SEQ ID NO: 21. 15. The kit of claim 1, further comprising one or more primers sets of SEQ ID NOs: 111 and 112, SEQ ID NOs: 113 and 114, SEQ ID NOs: 115 and 116, SEQ ID NOs: 117 and 118, and SEQ ID NOs: 119 and 120. 16. The kit of claim 1, further comprising forward primers comprising SEQ ID NOs: 29-34, 38-40, 42-64, 78-83, 86-91, 96, 98, 100-109 and reverse primers comprising SEQ ID NOs: 35-37, 41, 65-77, 84, 85, 92-95, and 97. 17. A method for assaying for two or more rearrangements, two or more translocations or at least one rearrangement and at least one translocation selected from the group consisting of a V.sub.H-J.sub.H IGH rearrangement, a D.sub.H-J.sub.H IGH rearrangement, a V.sub.K-J.sub.K IGK rearrangement, a V.sub.K/intron-Kde IGK rearrangement, a V.lamda.-J.lamda. IGL rearrangement, a V.beta.-J.beta. TCRB rearrangement, a D.beta.-J.beta. TCRB rearrangement, a V.sub.Y-J.sub.Y TCRG rearrangement, a V.delta.-J.delta. TCRD rearrangement, a D.delta.-D.delta. TCRD rearrangement, a D.delta.-J.delta. TCRD rearrangement, a V.delta.-D.delta. TCRD rearrangement, a t(11;14) (BCL1-IGH) translocation and t(14;18) (BCL2-IGH) translocation, using primers from the kit of claim 16 to amplify the corresponding locus in a sample. 18. A method according to claim 17 for the detection of minimal residual disease (MRD) or for identification of PCR targets to be used for MRD detection. 19. A method according to claim 18, wherein an amplified nucleic acid is detected using automated high resolution PCR fragment analysis. 20. The kit according to claim 16, further comprising primers comprising SEQ ID NOs: 111-120. 21. A method for detecting a D.sub.H-J.sub.H IGH rearrangement, comprising using the kit according to claim 1 to amplify the IGH locus in a sample. 22. The method of claim 21, wherein said IGH locus is amplified in a PCR assay. 23. The method of claim 22, wherein said PCR assay is a multiplex PCR assay. 24. A method for the detection of minimal residual disease (MRD) or for identification of PCR targets to be used for MRD detection comprising utilizing the primers of the kit of claim 1 to amplify nucleic acid in a sample. 25. A method for detecting a V.sub.H-J.sub.H IGH rearrangement, comprising using a primer kit comprising a plurality of forward primers selected from the group consisting of SEQ ID NOs: 1-8, 10-15, 17-20, and 22-28, and at least one reverse primer comprising SEQ ID NO:21 to amplify the IGH locus in a sample; wherein at least one of said plurality of forward primers or at least one of said at least one reverse primer comprises a fluorescent label.

Details for Patent 8,859,748

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2039-02-26