US 11,905,560 B2
Methods for multiplex detection of alleles associated with corneal dystrophy
Connie Chao-Shern, Menlo Park, CA (US)
Assigned to Avellino Lab USA, Inc.
Appl. No. 16/604,310
Filed by Avellino Lab USA, Inc., Menlo Park, CA (US)
PCT Filed Apr. 10, 2018, PCT No. PCT/US2018/026962
§ 371(c)(1), (2) Date Oct. 10, 2019,
PCT Pub. No. WO2018/191304, PCT Pub. Date Oct. 18, 2018.
Claims priority of provisional application 62/624,660, filed on Jan. 31, 2018.
Claims priority of provisional application 62/624,661, filed on Jan. 31, 2018.
Claims priority of provisional application 62/573,537, filed on Oct. 17, 2017.
Claims priority of provisional application 62/483,588, filed on Apr. 10, 2017.
Prior Publication US 2020/0040397 A1, Feb. 6, 2020
Int. Cl. C12Q 1/6883 (2018.01); C12Q 1/6827 (2018.01); G01N 33/58 (2006.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/6827 (2013.01); G01N 33/582 (2013.01); C12Q 2600/156 (2013.01); C12Q 2600/16 (2013.01)] 4 Claims
OG exemplary drawing
 
1. A method for detecting corneal dystrophy (CD) comprising:
(A-1) amplifying one or two DNA regions of interest from a biological sample from a subject using a reaction mixture comprising at least a first amplification primer pair to produce one or two amplified DNA regions of interest;
(B-1) hybridizing a first labeled TGFBI G623D mutant detection probe comprising the nucleotide sequence of SEQ ID NO: 36 and a second labeled TGFBI M502V mutant detection probe comprising the nucleotide sequence of SEQ ID NO: 30 to a first TGFBI gene sequence comprising a region encoding amino acid position 623 in the one or two amplified DNA regions of interest and a second TGFBI gene sequence comprising a region encoding amino acid position 502 in the one or two amplified DNA regions of interest, respectively;
and
(C) detecting CD if the at least G623D and M502V mutations in the one or two amplified DNA regions of interest are detected.