Multiplex PCR for detection of and in cattle.

genus has been classified under the family Anaplasmataceae of order Rickettsiales, which contains the spp. , , and is accountable for a broad range of diseases in both man and animal medicine around the globe. A multiplex PCR procedure was designed to identify , , and simultaneously by employing species-specific primers targeting the 4 (), 16S rRNA (), and () genes. The sensitivity of the assay was ascertained by tenfold dilutions of DNA obtained from cattle blood infected with , , and was used to evaluate the test sensitivity. A total of 31 genus positive samples were subjected to mPCR by using species specific primers for simultaneous detection of all the three species. All the 31 samples showed amplification of 753 bp fragment specific for and 61.29% of the samples showed amplification of 547 bp fragment specific for . The 470 bp DNA fragment specific for was found in 96.7% of samples. The lower limit of detection of the method for and was found to be 4.4, 0.44 and 0.044 ng/µl, respectively.