Cloning and sequencing of the ompA and smpA virulence genes of Acentobacter baumanniiisolated from clinical samples

Acinetobacterbaumannii has emerged as a medically important pathogen because of the increasing number of infections produced by this organism over the preceding three decades and the global spread of strains with resistance to multiple antibiotic classes. So, aim of this research, amplification, cloning and sequencing two virulence factor genes Acinetobacterbaumannii isolated from patients.
Collecting samples of Acinetobacterbaumannii taken from different clinical cases of wounds, septicemia, and urinary tract infections. That was accomplished by taking (30) samples from Imam Ali and Kashani hospitals Shahrekord Township. Samples were cultured on solid media (McConkey and blood agars), and according to microscopical, cultural, and biochemical were identified. The coding sequence of AcinetobacterbaumanniiompA and smpA genes was isolated by PCR method. The ompA and smpA genes was inserted into pTZ57R/T as T/A cloningvector. Transformation was confirmed with plasmid extraction, followed by double digestion and PCR methods.
A. baumannii isolates were identified in 10 different patients. All isolates (33.33%) were recovered from patients in the intensive care unit (ICU). As a result of PCR and double digestion two band 1150 and 411bp ompA and smpA genes respectively were observed. The sequences was found to be 90-95% similar to that ref sequences obtained in GenBank. The sequence of ompA and smpA genes amplified by the specific primer is closely matched (90 and 95% respectively) with aA. baumannii strains.
Transformation experiments revealed that these plasmids were capable to transform E. coli NB, an observation which indicates the ability of these plasmids to easy carrier large sequence into host. The ompA and smpA genes had “perfect” match (similarity, 90 and 95% respectively) with sequences of their corresponding gene (ompA and smpA genes) from GenBank as determined by using BLAST. So of this genes can used to construct DNA vaccine against Acinetobacterbaumannii.