Assay for human DNA for gender determination

• US 20060199217A1
• Filed: 05/10/2006
• Published: 09/07/2006
• Est. Priority Date: 09/30/2003
• Status: Active Grant

• Current Assignee: Reliagene Technologies Inc. (Laboratory Corporation Of America Holdings), Life Genetics Lab, LLC, Board of Supervisors of Louisiana State University Agricultural & Mechanical College (Louisiana State University System)
• Original Assignee: Life Genetics Lab, LLC
• Inventors: Batzer, Mark A., Sinha, Sudhir K., Hedges, Dale J.
• Granted Patent: US 7,432,362 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/6879 for sex determination
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: Y10S 435/975 Kit

Abstract

A method for determining gender from a human DNA sample. The loci of Alu element insertion is selected, amplified and evaluated in terms of size of the fragment. The gender assay utilizes AluSTXa for the X chromosome, AluSTYa for the Y chromosome, or both AluSTXa and AluSTYa, to reduce the possibility of error to a negligible quantity. The inserted chromosome yields a large fragment when the homologous region is amplified. The males are distinguished as having two DNA amplicons present, while females have only a single amplicon. The kit adapted for carrying out the method includes a pair of primers to amplify the locus and optionally polymerase chain reaction regents.

Claims

1. A primer pair for a sex determination of a human DNA sample, said primer pair comprising:

a forward primer and a backward primer, said primer capable of amplifying a selected locus of a human DNA in an amplification reaction, said selected locus being in a non-combining X—

Y homologous region, said region containing a monomorphic Alu insertion in one of the X chromosome and the Y chromosome.

2. The primer pair of claim 1, wherein said selected locus comprises STYa locus.

3. The primer pair of claim 2, wherein each of the primer pair has the sequence selected from, or constituting a subset of, the group consisting of:

4. The primer pair of claim 1, wherein said selected locus comprises STXa locus.

5. The primer pair of claim 4, wherein each of the primer pair has the sequence selected from, or constituting a subset of, the group consisting of:

6. A primer pair for a sex determination of a human DNA sample, said primer pair comprising:

a forward primer and a backward primer, said primer pair capable of amplifying alleles of Y-90 locus in an amplification reaction.

7. The primer pair of claim 6, wherein each primer of the primer pair has the sequence selected from, or constituting a subset of, the group consisting of:

8. A kit for determining gender from a sample, comprising:

a polymerase chain reaction reagent; and

a pair of primers capable of amplifying a selected locus of a human DNA in an amplification reaction, said selected locus being in a non-combining X-Y homologous region, said region containing a monomorphic Alu insertion in one of the X chromosome and the Y chromosome.

9. The kit of claim 8, wherein the selected locus is AluSTYa locus on the Y chromosome.

10. The kit of claim 9, wherein each primer of the primer pair has the sequence selected from, or constituting a subset of, the group consisting of:

11. The kit of claim 8, wherein the selected locus is AluSTXa locus on the X chromosome.

12. The kit of claim 11, wherein each primer of the primer pair has the sequence selected from, or constituting a subset of, the group consisting of:

13. The kit of claim 8, wherein the selected locus comprises AluSTYa locus and AluSTXa locus.

14. The kit of claim 13, wherein said pair of primers comprises:

a first primer pair, each primer of the first primer pair having the sequence selected from, or constituting a subset of, the group consisting of;

(SEQ ID NO;

     3)Forward 5′

-TGAAGAAATTCAGTTCATAGCTTGT-3′

;

and (SEQ ID NO;

     4)Reverse 5′

-CAGGAGATCCTGAGATTATGTGG-3′

;

and

a second primer pair, each primer of the second primer pair having the sequence selected from, or constituting a subset of, the group consisting of;

(SEQ ID NO;

     1)Forward 5′

-ACTGCAGTCCGCAGTCCGG-3′

;

and (SEQ ID NO;

     2)Reverse 5′

-CCACTGACATATTATCAGCCTTATTAC-3′

.

15. A kit for determining gender from a sample, comprising:

a polymerase chain reaction reagent; and

a pair of primers capable of amplifying alleles of Y-90 locus in an amplification reaction.

16. The kit of claim 15, wherein each primer of said pair of primers contains the sequence selected from, or constituting a subset of, the group consisting of: