Method of detecting and predicting bronchodilatory response to beta agonist
View Patent ↗Present invention relates to a method for predicting an individual's bronchodilatory response to a β agonist. Present invention particularly relates to the detection of specific allelic variants of the β2AR gene and their use as pharmacogenetic markers towards response to β agonist.
1. A method for detecting a single nucleotide polymorphism A46G in a β2AR gene in a human subject suffering from asthma and identifying the human subject as a good responder to a short acting β2 agonist based on the genotype at nucleotide position 46 of the beta 2AR gene said method comprising the steps of:
(a) isolating genomic DNA from the human subject,
(b) amplifying the genomic DNA using oligonucleotide PCR primers set forth in SEQ ID NOS. 2 and 3,
(c) sequencing the PCR amplified genomic DNA obtained in step (b)
(d) detecting the single nucleotide polymorphism A46G in the sequenced PCR amplified genomic DNA from step (c),
(e) identifying specific β2AR genotypes AA, AG and GG, and
(f) based on the information from step (e), identifying the human subject to be a good responder to the short acting β2 agonist if said subject has GG genotype and a poor responder to the short acting β2 agonist if said subject has the AA genotype.
2. A method as claimed in claim 1 , wherein the short acting β2-agonist is salbutamol.
3. A method as claimed in claim 2 , wherein the pharmacologically active dose of β2-agonist, salbutamol, is in the range of 100 to 250 μg.
4. The method of claim 1 wherein said detecting comprises amplification and/or primer extension with primers selected from the group consisting of:
(a) oligonucleotide primers having SEQ ID Nos. 2 and 3,
(b) oligonucleotide primers having SEQ ID Nos. 4 and 5, and
(c) oligonucleotide primers having SEQ ID Nos. 6,7,8 and 9.
5. The method of claim 4 , wherein said detecting comprises:
(a) denaturing the isolated DNA at temperature of 96° C. for 10 seconds,
(b) annealing the primers of SEQ ID No. 4 and 5 to denatured DNA of step (a) at a temperature of 55° C. for 5 seconds, and
(c) undertaking the extension of annealed DNA of step (b) at a temperature of 60° C. for 30 seconds.
6. The method of claim 5 , further comprising amplification and/or primer extension from primers having SEQ ID Nos. 6, 7, 8 and 9.