Methods for predicting increase sperm ejaculate in a varicocele associated fertility patient
View Patent ↗The present invention provides a method for predicting whether a patient having varicocele-associated infertility will have an increase in sperm in the ejaculate after varicocele repair surgery. In the method of the present invention, the present or absence of undelete forms of exons 6, 7, 8 and 9 the patient's L-VDCC α1c transcript is detected. The presence of undeleted forms of the exons indicates that varicocele repair surgery will result in increased ejaculate, and the absence of undeleted forms of the exons indicates that varicocele repair surgery will not result in increased ejaculate.
1. A method for predicting whether a patient having varicocele-associated infertility will have an increase in sperm in the ejaculate after varicocele repair surgery, said method comprising detecting in the patient the presence or absence of an undeleted form of an L-VDCC α1c transcript, where the presence of an undeleted form of the L-VDCC α1c transcript indicates that varicocele repair surgery will result in a greater than 50% increase in sperm in the ejaculate after varicocele repair surgery, and the absence of an undeleted form of the L-VDCC α1c transcript predicts that varicocele repair surgery will not result in an increased sperm count in the ejaculate after varicocele repair surgery.
2. The method of claim 1 , comprising:
obtaining a suitable tissue sample from the patient;
preparing from the tissue sample a nucleic acid sample; and
analyzing the nucleic acid sample for the presence or absence of said undeleted form of said transcript.
3. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:7.
4. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:8.
5. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:9.
6. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:10.
7. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:11.
8. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:12.
9. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:13.
10. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:14.
11. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:15.
12. The method of claim 2 , wherein said analyzing is performed using an oligonucleotide probe having the sequence of SEQ ID NO:16.
13. The method of claim 2 , wherein said analyzing is performed using a temperature that is at or less than 10° C. below the melting temperature of a perfect hybrid formed between a probe and said transcript.
14. The method of claim 2 , wherein said analyzing is performed between 42° C. and 60° C.
15. The method of claim 2 , wherein prior to said analyzing, said transcript is amplified.
16. The method of claim 15 , wherein said transcript is amplified by PCR.
17. The method of claim 16 , wherein the PCR amplification is carried out using at least one oligonucleotide primer selected from the group of primers consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.
18. The method of claim 16 , wherein the PCR amplification is carried out using at least one forward primer and at least one reverse primer.
19. The method of claim 2 , wherein the tissue sample is a testis biopsy.