The study was undertaken with an objective to evaluate a rapid PCR based method for detection of adulteration of buffalo milk in cow milk at minimum level of detection. This method utilizes primers targeting the mitochondrial encoded 12S rRNA gene as the target for species identification. PCR assay involve use of three different primers. Reverse primers specific for cow and buffalo complementary to the gene fragment of 12S rRNA along with the common forward primer. The cow specific primer, along with the common forward primer, yields a cow specific amplicon of 346 bp in the 12S rRNA gene. On the other hand, a buffalo specific primer along with the same common forward primer yields a buffalo specific amplicon of 220 bp fragment in the same gene. The method evaluated was able to detect presence of buffalo milk in cow at level of adulteration.
