Isolation, Identification And Sequencing Of Mycoplasma Galisepticum By Culture And PCR In Baghdad City, Iraq

This study is carried out to investigate the presence of Mycoplasma galisepticum in broiler and layer in farms infected with respiratory signs, a total of 200 samples were collected randomly from layer and broiler chickens with respiratory signs in Baghdad between January to May 2017. the results of culture revealed that the total rate of Mycoplasma species isolates (positive cultures) was 10%. The highest number of Mycoplasma species isolates was seen in trachea and lowest in conjuctivial swabs, oviduct and lungThere was no isolate from the choanal and nasal swabs, all positive cultures were staining with Diene's stain. DNA was extracted from 20 positive culture isolates, all twenty isolates were positive for Mycoplasma genous by convential PCR assay, and a product of 250 base pair was generated by amplification of 16S rRNA gene. Then twenty positive Mycoplasma spp isolates were subjected to MG and MS detection Kit, the result revealed 2/20 (10%) MG, 11/20 (55%) MS and 7/20 (35%) were positive for both MG and MS. The two MG isolates confirmation by subjected to amplification of the 16S RNA gene the results revealed that MG specific primers of the16S RNA gene the single band at 183bp the product of amplification of Mycoplasma galisepticum 16SrRNA gene was sent by MACROGEM (Korea) for sequencing.were submitted in Gene bank database and have accession. number: ID: MG846120.1 Sequencing alignment showed that the 99% similarity of first isolate and second isolate of MG field isolates with standard reference Mycoplasma galisepticum strain ATCC, the phylogentic analysis revealed that it was the Iraqi isolate was 100% similar to USA strain1NR104952, south Africa strain2MF196172, Iran strain7KC865737, VietNamHatey strain 3AMO75207 and Egypt strain 6GO902040. Also which had 99% similar to Russia strain 8L36043.3, India strain 4KP685378.1 and Iran strain5AY705443.1.