Optimization of polymerase chain reaction for direct detection of colibacillosis in infected chicken | ||
| Veterinary Medical Journal (Giza) | ||
| Volume 59, Issue 4, October 2011, Pages 307-318 PDF (1.79 M) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/vmjg.2011.368354 | ||
| Authors | ||
| El-Seedy R* 1; Hassan H1; Salama S2; Fatma El-Zahraa Gamal2 | ||
| 1Dept.of Bacteriology, Mycology and Immunology, Faculty of Vet. Med, Beni-Suef University | ||
| 2Central Laboratory for Evaluation of Veterinary Biologics | ||
| Abstract | ||
| A total of 33 local E. coli isolates were 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 16S-RNA, 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 and O78 types 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 type 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. | ||
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