Optimization of polymerase chain reaction for direct detection of colibacillosis in infected chickens

A total of 33 local E. coli isolates were used in this study. These isolates were biochemically and serologically identified as O1, O2, O6, O78 and O126. Four sets of oligonucleotide primer sequences were designed specifically for 16SrRNA, STX, Sth and eaeA genes. DNA and Plasmids were extracted and polymerase chain reaction was optimized for each. 16SrRNA gene primer successfully amplified with all serotypes giving rise a product mass of 204 bp, while STX gene primer was amplified with O1, O2 an dO78 serotypes in a specific band at 323 bp. At the same time the specific primers of Sth gene get a 171 bp molecular weight product only with O6 serotype meanwhile the eaeA gene primers successfully amplified only with O126 serotype giving rise a molecular weight band at 200 bp. In conclusion, PCR assay was able to differentiate between the different serotypes of E. coli in the suspected samples saving time, money and effort

Evaluation of the recombinant m9 protein as haemagglutinating antigen for serodiagnosis of M. gallisepticum in chickens

This study focused on a prominent haemagglutination feature of the pathogenic M. gallisepticum, which have agglutinins that are immunogemc surface protein. M9 protein was prepared using cloning and expression technology and evaluated as haemagglutinating antigen. In comparing its haemagglutination properties with the whole cell antigen, the M9 protein showed somewhat less sensitivity starting after the first week post infection with M. gallisepticum strainsand reaching the maximum reactivity at the seventh week post infection, while the whole cell antigen reached its maximum reactivity at the sixth week post infection. Regarding the specificity, both antigens are highly specific and showed no reactions with the M. synoviae antisera. The advantages of M9 protein as haemagglutinating antigen is that it could be used in different areas and is cheaper in preparation

Evaluation of multiplex pcr based test in comparison with culture method for detection of mycoplasmosis in infected chickens

A total of 94 chicken samples were examinedfor Mycoplasma gallisepticum and Mycoplasma syjzoviae either by culturing procedure or by using the multiplex PCR. The comparative analysis of the obtained results indicated that the multiplex PCR was more sensitive and reliable in the detection of both organisms, where the multiplex PCR detected Mycoplasma gallisepticum in 20.21% of the tested samples while it gave 5.319% prevalence forMycoplasma synoviae. Moreover isolation and culturing procedures detected Mycoplasma gallisepticum and Mycoplasma synoviae in a prevalence rate of 15.956% and 2.127% of the tested samples comparatively. So these results concluded that, the multiplex PCR has become a valuable and easy test aiding in diagnosis of M. gallisepticum and M. synoviae infection in chickens