Genetic test for detecting congenital adrenal hyperplasia

Abstract

A method for amplifying a CYP21A2 gene and/or a CYP21A2 gene chimera from a sample is provided. In some embodiments, the method may comprise amplifying a product from a sample comprising human genomic DNA by PCR using a forward primer that is complementary to a sequence that is duplicated in a bimodular human RCCX locus and a reverse primer that is complementary to a sequence that occurs only once in the bimodular human RCCX locus at a position that is downstream of the CYP21A2 gene. Methods for analyzing the amplification product are also provided.

1 - 14 . (canceled) 
     
     
         15 . A set of reagents comprising:
 (a) a forward primer that is complementary to a sequence that is duplicated in a bimodular human RCCX locus, wherein the bimodular human RCCX locus comprises, in order, an RP1 gene, a C4A gene, a CYP21A1P pseudogene, a TNXA gene, an RP2 gene, a C4B gene, a CYP21A2 gene and a TNXB gene, and:
 i. the first occurrence of the duplicated sequence is in the C4A gene and/or the RP1 gene; and 
 ii. the second occurrence of the duplicated sequence is in the C4B gene and/or the RP2 gene; 
   (b) a reverse primer that is complementary to a sequence that occurs only once in the bimodular human RCCX locus at a position in the TNXB gene;   (c) a first primer mix and a second primer mix, wherein:
 i. the first primer mix comprises multiple primer pairs that target multiple non-overlapping segments that are in both the CYP21A2 gene and the CYP21A1P pseudogene, and 
 ii. the second primer mix comprises multiple primer pairs that target multiple non-overlapping segments that are in both the CYP21A2 gene and the CYP21A1P pseudogene; and 
 wherein at least some of the segments targeted by the first primer mix overlap with the segments targeted by the second primer mix. 
   
     
     
         16 . The set of reagents of  claim 15 , wherein the first primer mix and second primer mix each independently comprise 5-15 primer pairs. 
     
     
         17 . The set of reagents of  claim 15 , wherein the primers of (c) are indexed. 
     
     
         18 . The set of reagents of  claim 15 , wherein the primers of (c) are independently in the range of 15 to 50 nucleotides in length. 
     
     
         19 . The set of reagents of  claim 15 , wherein the median length of the segments of (c) is in the range of 100 by to 1 kb. 
     
     
         20 . The set of reagents of  claim 15 , wherein the median length of the segments of (c) is in the range of 150 bp to 500 bp. 
     
     
         21 . The set of reagents of  claim 15 , wherein the segments of (c) comprise all 10 exons of the CYP21A2 gene.