Molecular method for universal detection of citrus viroids

Patent metadata

Field	Value
Publication number	US-9574244-B2
Application number	US-201414341581-A
Country	US
Kind code	B2
Filing date	Jul 25, 2014
Priority date	Nov 7, 2011
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

The present invention provides methods for universally detecting citrus viroids in plant material such as germplasm. In particular embodiments, the invention enables the determination of citrus viroid infection and plant resistance. Accordingly, the present method provides methods for improved universal detection of any citrus viroid.

First claim

What is claimed is:

1. A diagnostic reaction mixture composition comprising a nucleic acid sample, a degenerate primer pair comprising SEQ ID NOS: 1 and 2, and/or a degenerate primer pair comprising SEQ ID NOS: 3 and 4.

2. The composition of claim 1 , wherein said degenerate primer pair comprises SEQ ID NOS: 1 and 2.

3. The composition of claim 1 , wherein said degenerate primer pair comprises SEQ ID NOS: 3 and 4.

4. The composition of claim 1 , wherein said degenerate primer pair comprises SEQ ID NOS: 1 and 2, and wherein the mixture further comprises a second primer pair comprising SEQ ID NOS: 3 and 4.

5. The composition of claim 1 , wherein said nucleic acid sample is extracted from plant material.

6. The composition of claim 1 , further comprising dNTPs, a buffer, DNA polymerase, reverse transcriptase, a fluorophore, or a stabilizer.

7. A kit comprising one or more a degenerate primer pair comprising SEQ ID NOS: 1 and 2 and/or SEQ ID NOS: 3 and 4.

8. The composition of claim 5 , wherein said plant material is selected from a group consisting of seed, foliage, limbs, trunk, bark, rootstock, fruit, germplasm, propagule, cuttings, and budwood.

9. The composition of claim 1 , further comprising a fluorescent dye.

10. The composition of claim 1 , wherein at least one primer of the degenerate primer pair is coupled to a fluorophore.

11. The kit of claim 7 , wherein said degenerate primer pair comprises SEQ ID NOS: 1 and 2.

12. The kit of claim 7 , wherein said degenerate primer pair comprises SEQ ID NOS: 3 and 4.

13. The kit of claim 7 , wherein said degenerate primer pair comprises SEQ ID NOS: 1 and 2, and wherein the kit further comprises a second primer pair comprising SEQ ID NOS: 3 and 4.

14. The kit of claim 7 , further comprising dNTPs, a buffer, DNA polymerase, reverse transcriptase, a fluorophore, or a stabilizer.

15. The kit of claim 7 , further comprising a fluorescent dye.

16. The kit of claim 7 , wherein at least one primer of the degenerate primer pair is coupled to a fluorophore.

Assignees

• Univ California

Inventors

• Vidalakis Georgios
• Wang Jinbo

Classifications

• C12Q2600/112

  Disease subtyping, staging or classification · CPC title

• C12Q1/6895

  for plants, fungi or algae · CPC title

• C12Q2600/158

  Expression markers · CPC title

• C12Q1/701 Primary

  Specific hybridization probes · CPC title

• C12Q2600/16

  Primer sets for multiplex assays · CPC title