Abstract：The genotyping system based on short tandem repeats (STR) and sex locus can not only provide important molecular genetic marker information, but also effectively solve the problem of individual identification and genetic relationship in animals. To establish a multiplex detection system for STR and CHD (chromobox-helicase-DNA binding gene) of pigeon (Columba livia domestica) and evaluate its forensic application value, in present study, a 6-dye fluorescence-labelling multiplex PCR amplification system for 19 STR (short tandem repeats) loci and 1 CHD sex locus was established to evaluate its forensic application and population research. In total, CliμD11, PG4, PG1, PG2, CliμT02, CliμD17, CliμD35, CliμT17, CliμD16, PIGN04, CliμD32, PIGN57, PIGN26, PG5, PG6, CliμD19, PIGN12, PIGN15, PIGN10, and gender-identification gene CHD were picked up. The results showed that the multiplex amplification system was reliable and easy for identification via feather examples from 241 pigeons. It had been proved that the sensitivity could be low to 0.0625 ng DNA and the species identification was rather specific. The statistic analysis of all alleles of 19 STR loci in the pigeon population showed that all 19 STR loci could be qualified to be used in individual identification and paternity test. Finally it was proved that the pigeon STR loci multiplex amplification system established in present research was rather stable, sensitive and species-specific, which was the most effective pigeon detection system and the first report in China as well. The present study research provides molecular basis for individual identification and paternity testing in pigeons.
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