Molecular Identification of Acinetobacter baumannii Isolated from Clinical Samples in Iraq by Using Multiplex PCR

Authors

Yaseen Ismael Mamoori, Department of Molecular and Medical Biotechnology, College of Biotechnology, Al-Nahrain University, Jadriya Baghdad, Iraq.
Ibrahim Abdulla Ahmed, Department of Plant Biotechnology, College of Biotechnology, Al-Nahrain University, Jadriya Baghdad, Iraq.
May Ridha Jaafer, Department of Gene Bank, Forensic DNA Center for Research and Training, Al-Nahrain University, Jadriya, Baghdad, Iraq.
Ayat Muneam Ali, Department of Applied Pathological Analysis, College of Science, Al-Nahrain University, Jadriya, Baghdad, Iraq.
Sabreen Ali Hadi, Department of Plant Biotechnology, College of Biotechnology, Al-Nahrain University, Jadriya Baghdad, Iraq.

Abstract

Acinetobacter baumannii is one of the most clinically significant pathogens that cause severe nosocomial infections. Rapid identification of this bacterial species is essential so that proper treatment can be provided and outbreaks can be controlled. In this study, a multiplex PCR was used to identify A. baumannii. Twenty clinical isolates of A. baumannii and three each of Gram+ve (Staphylococcus aureus, Bacillus licheniformis, Micrococcus yunnanensis) and Gram−ve (Proteus mirabilis, Pseudomonas aeruginosa, Klebsiella pneumoniae) bacterial isolates were subjected to singleplex and multiplex PCR reactions using three sets of species-specific primers (P-Ab-ITS-F, P-Ab-ITS-R; OXA-51-like-F, OXA-51-like-R; sp2-F, sp4-R). Additionally, 24 Acinetobacter spp., including A. baumannii genomes, were retrieved from the National Center for Biotechnology Information GenBank database and aligned with each primer using the Blast program. The complementary bases of the primers with the bacterial genome were determined, and their melting temperatures were calculated using the Promega Tm calculator. Multiplex PCR successfully amplified three amplicons in all the tested A. baumannii isolates; however, it did not amplify other bacterial species. Furthermore, in comparison to other Acinetobacter spp., there were significant differences in the mean annealing temperatures of the primer sets for A. baumannii (pA. baumannii. Moreover, it is feasible in most laboratories and is essential for clinicians, the control of outbreaks, and epidemiological studies.

Keywords

Acinetobacter baumannii, blaOXA-51-like gene, gyrB gene, intergenic spacer (ITS) region, multiplex PCR

Subject Area

Biology

Article Type

Article

First Page

2619

Last Page

2628

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite this Article

Mamoori, Yaseen Ismael; Ahmed, Ibrahim Abdulla; Jaafer, May Ridha; Ali, Ayat Muneam; and Hadi, Sabreen Ali (2025) "Molecular Identification of Acinetobacter baumannii Isolated from Clinical Samples in Iraq by Using Multiplex PCR," Baghdad Science Journal: Vol. 22: Iss. 8, Article 14.
DOI: https://doi.org/10.21123/2411-7986.5025