US 11,655,509 B2
Kits for diagnosing spinal muscular atrophy and uses thereof
Yi-Yi Kuo, New Taipei (TW); I-Fan Chiu, Hsinchu (TW); Lai-Ha Chung, New Taipei (TW); and Shu-Ju Lee, New Taipei (TW)
Assigned to Origin Biotechnology Co., Ltd., Taipei (TW)
Filed by Origin Biotechnology Co., Ltd., Taipei (TW)
Filed on Jan. 18, 2021, as Appl. No. 17/151,225.
Claims priority of application No. 202010138478.6 (CN), filed on Mar. 3, 2020.
Prior Publication US 2021/0277473 A1, Sep. 9, 2021
Int. Cl. C12Q 1/6883 (2018.01)
CPC C12Q 1/6883 (2013.01) [C12Q 2600/156 (2013.01); C12Q 2600/158 (2013.01); C12Q 2600/16 (2013.01)] 4 Claims
 
1. A kit for making a diagnosis as to whether a human subject has spinal muscular atrophy (SMA) or is a carrier of SMA, comprising,
a first primer consisting of the polynucleotide sequence of SEQ ID NO: 9;
a second primer consisting of the polynucleotide sequence of SEQ ID NO: 10;
a third primer consisting of the polynucleotide sequence of SEQ ID NO: 11;
a fourth primer consisting of the polynucleotide sequence of SEQ ID NO: 12;
a fifth primer consisting of the polynucleotide sequence of SEQ ID NO: 13;
a sixth primer consisting of the polynucleotide sequence of SEQ ID NO: 14;
a seventh primer consisting of the polynucleotide sequence of SEQ ID NO: 7;
an eighth primer consisting of the polynucleotide sequence of SEQ ID NO: 8; and
polymerase chain reaction (PCR) reagents.
 
4. A method of making a diagnosis as to whether a human subject has spinal muscular atrophy (SMA) or is a carrier of SMA by using the kit of claim 1, comprising,
(a) extracting a DNA sample from the human subject;
(b) mixing the DNA sample with the first to eighth primers in the PCR reagents;
(c) subjecting the mixture of the step (b) to PCR;
(d) determining a first copy number of exon 7 of survival of motor neuron 1 (SMN1) gene, a second copy number of exon 7 of survival of motor neuron 2 (SMN2) gene, a third copy number of exon 8 of the SMN1 gene, and a fourth copy number of exon 8 of the SMN2 gene in the DNA sample from the product of the step (c); wherein when the sum of the first and the second copy numbers is not equal to the sum of the third and fourth copy numbers, then repeating steps (b) and (c) until the sum of the first and the second copy numbers equals to the sum of the third and fourth copy numbers; and
(e) making the diagnosis of SMA based on the first copy number determined by the step (d), wherein
when the first copy number is 0, then diagnosing the human subject as having SMA; or
when the first copy number is 1, then diagnosing the human subject as being a carrier of SMA.