Vet World Vol.11 July-2018 Article-19
Research Article
Veterinary World, 11(7): 1006-1014
https://doi.org/10.14202/vetworld.2018.1006-1014
Use of molecular biology tools for rapid identification and characterization of Pasteurella spp.
2. Aerobic Bacterial Vaccine Research Department, Veterinary Serum and Vaccine Research Institute, Cairo Egypt.
3. Anaerobic Bacterial Vaccine Research Department, Veterinary Serum and Vaccine Research Institute, Cairo Egypt.
4. Department of Microbiology and Immunology, Veterinary Division, National Research Center, Dokki, Egypt.
Background and Aim: This study aimed to create rapid characterization and genotyping of Pasteurella multocida (PM) protocol using modern molecular biology techniques.
Materials and Methods: Thirty bacterial isolates were characterized by capsular and somatic identification using conventional procedure followed by multiplex polymerase chain reaction (PCR), restriction endonucleases analysis (REA), and finally confirmed by sequence analysis. Two local vaccine strains and two field isolates were identified as PM Type A and B.
Results: A total of 30 isolates were found positive for PM either morphologically and biochemically; however, multiplex PCR technique identified only 22 isolates as Pasteurella species using universal primers while 8 isolates were found negative for PM. 12 of 22 isolates (54%) were characterized at the same reaction into PM Type A, five isolates (23%) were Type B and the rest five isolates (23%) of tested isolates were negative for Types A, B, and D. Hemorrhagic septicemia Type B: 2 or B: 5 could be identified somatically within PM capsular serogroup B using PCR technique. Somatic characterization of PM was done using REA that could identify all PM Type A into A:1 and all PM Type B into B: 2. These protocols were verified for its accuracy and reliability by sequence analysis of two vaccine strains of PM Type A and B that were characterized previously by biochemical and serological methods as well as two selected isolates from the 22 positive isolates representing PM Type A and B.
Conclusion: PCR and REA could confirm the identity of PM and provide a rapid and reliable characterization in comparison with biochemical analysis and conventional serotyping that may take up to 2 weeks. Hence, they can reduce the time needed for polyvalent vaccine production and when the reference antisera are unavailable. Moreover, the identity of Omp-H for vaccine and field strains may provide better data to control Pasteurellosis in Egypt. Keywords: multiplex polymerase chain reaction, outer membrane protein H, Pasteurella multocida, restriction endonucleases analysis.
Keywords: multiplex polymerase chain reaction, outer membrane protein H, Pasteurella multocida, restriction endonucleases analysis.
How to cite this article: Abbas AM, Abd El-Moaty DAM, Zaki ESA, El-Sergany EF, El-Sebay NA, Fadl HA, Samy AA (2018) Use of molecular biology tools for rapid identification and characterization of Pasteurella spp., Veterinary World, 11(7): 1006-1014.
Received: 15-02-2018 Accepted: 07-06-2018 Published online: 29-07-2018
Corresponding author: Dalia A. M. Abd El-Moaty E-mail: dody.ahmed@gmail.com
DOI: 10.14202/vetworld.2018.1006-1014
Copyright: Abbas, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.