The present study has investigated on ALH class II -DQB1 gene in population of Srirangapatna at the southern region of India. 42 individuals in comparison with their most homologous alleles have shown new nucleotide substitutions which lead to new alleles. 22.2% of new alleles comprised only one or two nucleotide variations. DQB1*03 is the most frequent allele group in the present study which has displayed by 20 (44.5%) samples and allele group DQB1*02 with six samples (13.33%) as the lowest frequency. Allele DQB1*03:23:01 was demonstrated as most frequent allele by 18 (40%) samples. We identified 87% of alleles represent at least one non-synonymous mutation while 6% of alleles having only synonymous nucleotide variations. According to numerous ethnicities in India, high-resolution HLA typing is essential to figure out new HLA alleles which have not detected so far through classical approaches.