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Development of multiplex PCR assay for species identification of cow, pig,
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chicken and duck from raw meats
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Abstract :
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A multiplex polymerase chain reaction(PCR) assay was developed for the simultaneous
identification of four species of cow, pig, chicken and duck from raw meats. The
primers used in this study were designed in different regions of mitochondrial DNA(16S
RNA) after alignment of the available sequences in the Genbank database. The primers
generated specific fragments of 91 bp, 181 bp, 279 bp and 477 bp lengths for pig,
chicken, cow and duck, respectively. The detection limit of the multiplex PCR assay
ranged between 1 pg and 0.1 pg of template DNA from extracted from four raw
meats. Enzyme linked immunosorbent assay(ELISA), a commercially available meat
identification kit, was performed to compare with multiplex PCR assay. Total sixty six
samples(cow 32, pig 12, chicken 12, and duck 10) were tested for comparison
between PCR assay and ELISA kit. The species specificity was 100% in all four species
in the multiplex PCR assay, In contrast, the species specificity of ELISA kit were 100%,
100%, 90.6%, 83.3% in the chicken, duck, cow, and pig, respectively. In, addition,
when this ELISA kit and PCR test were applied to other animal species of horse,
sheep, goat and turkey, no amplification and negative reaction were shown in the PCR
and ELISA kit, respectively. These studies suggest that this multiplex PCR assay can be
used for rapid and simultaneous species identification of cow, pig, chicken and duck
from raw meats. Further studies are needed for evaluation of developed multiplex PCR
from various samples including meat products.
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