Primers for Diagnosing Avellino Corneal Dystrophy

Abstract

The present invention relates to a real-time PCR primer pair and probe for diagnosing Avellino corneal dystrophy, and more particularly to a real-time PCR primer pair and probe for diagnosing Avellino corneal dystrophy, which can accurately diagnose the presence or absence of a mutation in exon 4 of BIGH3 gene, which is responsible for Avellino corneal dystrophy. The use of the primer pair and probe according to the invention can diagnose Avellino corneal dystrophy in a more rapid and accurate manner than a conventional method that uses a DNA chip or PCR.

1 - 3 . (canceled) 
     
     
         4 . A real-time PCR reagent composition comprising:
 (i) a first primer having the sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, or 23; and   (ii) a second primer having the sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, or 24.   
     
     
         5 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 1, and a second primer having the sequence of SEQ ID NO: 2. 
     
     
         6 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 3, and a second primer having the sequence of SEQ ID NO: 4. 
     
     
         7 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 5, and a second primer having the sequence of SEQ ID NO: 6. 
     
     
         8 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 7, and a second primer having the sequence of SEQ ID NO: 8. 
     
     
         9 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 9, and a second primer having the sequence of SEQ ID NO: 10. 
     
     
         10 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 11, and a second primer having the sequence of SEQ ID NO: 12. 
     
     
         11 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 13, and a second primer having the sequence of SEQ ID NO: 14. 
     
     
         12 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 15, and a second primer having the sequence of SEQ ID NO: 16. 
     
     
         13 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 17, and a second primer having the sequence of SEQ ID NO: 18. 
     
     
         14 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 19, and a second primer having the sequence of SEQ ID NO: 20. 
     
     
         15 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 21, and a second primer having the sequence of SEQ ID NO: 22. 
     
     
         16 . The real-time PCR reagent composition of  claim 4  comprising a first primer having the sequence of SEQ ID NO: 23, and a second primer having the sequence of SEQ ID NO: 24. 
     
     
         17 . The real-time PCR reagent composition of  claim 4  further comprising:
 (i) a first probe having the sequence of SEQ ID NO: 25, 27, 29, 31, 33, 35, 37, 39, or 41; and 
 (ii) a second probe having the sequence of SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, or 42. 
 
     
     
         18 . The real-time PCR reagent composition of  claim 17  comprising:
 a first probe having the sequence of SEQ ID NO: 25 and a second probe having the sequence of SEQ ID NO: 26, 
 a first probe having the sequence of SEQ ID NO: 27 and a second probe having the sequence of SEQ ID NO: 28, 
 a first probe having the sequence of SEQ ID NO: 29 and a second probe having the sequence of SEQ ID NO: 30, 
 a first probe having the sequence of SEQ ID NO: 31 and a second probe having the sequence of SEQ ID NO: 32, 
 a first probe having the sequence of SEQ ID NO: 33 and a second probe having the sequence of SEQ ID NO: 34, 
 a first probe having the sequence of SEQ ID NO: 35 and a second probe having the sequence of SEQ ID NO: 36, 
 a first probe having the sequence of SEQ ID NO: 37 and a second probe having the sequence of SEQ ID NO: 38, 
 a first probe having the sequence of SEQ ID NO: 39 and a second probe having the sequence of SEQ ID NO: 40, or 
 a first probe having the sequence of SEQ ID NO: 41 and a second probe having the sequence of SEQ ID NO: 42. 
 
     
     
         19 . The real-time PCR reagent composition of  claim 17 , wherein the first real-time PCR probe comprises a first non-naturally occurring label and the second real-time PCR probe comprises a second non-naturally occurring label. 
     
     
         20 . The real-time PCR reagent composition of  claim 19 , wherein the first and/or second non-naturally occurring label is a fluorescent label. 
     
     
         21 . The real-time PCR reagent composition of  claim 19 , wherein the first and/or second non-naturally occurring label is either VIC or FAM. 
     
     
         22 . The real-time PCR reagent composition of  claim 19 , wherein the second probe further comprises a minor groove binder. 
     
     
         23 . A method of detecting Avellino corneal dystrophy, comprising detecting a mutation in the βIG-H3 gene by using the real-time PCR reagent composition of  claim 4 .