Molecular discrimination for the most pathogenic serotypes of clinical isolates of Klebsiella pneumoniae using cps locus encoded to capsular antigens. | ||||
| Microbes and Infectious Diseases | ||||
| Articles in Press, Accepted Manuscript, Available Online from 18 July 2025 | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/mid.2025.399945.2978 | ||||
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| Authors | ||||
Abdulameer M. Ghareeb   1; Tamara H. Zedan2; Fatimah A. Abdul Jabbar 2
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| 1Institute of Genetic Engineering and Biotechnology, University of Baghdad, Baghdad, Iraq | ||||
| 2College of Biotechnology, Al-Nahrain University, Baghdad, Iraq | ||||
| Abstract | ||||
| Abstract: Klebsiella pneumoniae predominant microorganisms implicated in Urinary tract infections (UTIs) that can be life-threatening. Molecular typing and virulence pattern analysis are required to discriminate bacteria. Objectives: This study aimed to molecular discrimination and genetic relatedness of multidrug resistant K. pneumoniae isolates with investigate the virulome pattern. Methods: Two hundred and seventy urine samples were collected from UTI patients (both sex age ranged 18-50 years) at three main hospitals in Baghdad. Isolates identified using conventional molecular methods. Isolates genotype was conducted to determine the presence of two target cps cluster (wzc, orf10) genes mediated biosynthesis of capsular polysaccharides in both types (K1 and K2) respectively. Antibiotics susceptibility test and occurrence of virulence encoded genes (fimH, mrkD, wabG) was determined among 20 selected isolates. Further discrimination was conducted utilizing Enterobacterial repetitive intergenic consensus ERIC-PCR technique on these isolates. Results: Sixty MDR K. pneumoniae isolates (22%) were isolated from urine samples. 85% of isolates showed ability to produce biofilm.The results confirmed the presence amplicon of 16S rRNA with expected size (130bp) in all tested isolates. Examined isolates were divided to 29 (48.3%) isolates were (K1) capsular serotype according to expected amplicon size 350bp for wzc gene while only 20 (33.3%) isolates showed a band with size of 650bp for orf10 and consider as a K2 serotype in addition to 11 (18.3%) isolates were neither K1 nor K2 (non-typeable by K1 and K2). The PCR detection showed that 20 (100%), 17 (85%), and 16(80%) of isolates carried fimH, mrkA, and wabG, respectively. ERIC-PCR analysis grouped the isolates into 4 clusters according to their profile amplicons. Conclusion: Both wzc and orf10 genes are reliable marker to identify K. pneumoniae serotypes K1 and K2. Although there is high prevalence of biofilm encoded genes, there was no significant difference in prevalence rates among three groups of serotypes. ERIC genotyping methods had no correlation with the capsule types. | ||||
| Keywords | ||||
| capsular polysaccharides; Klebsiella pneumoniae; mrkD protein; ERIC; fimbria | ||||
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