Prevalence of pathogenic Klebsiella pneumoniae based on PCR capsular typing harbouring carbapenemases encoding genes in Uganda tertiary hospitals

Abstract

Background Klebsiella pneumoniae is an opportunistic pathogen that has been implicated as one of commonest cause of hospital and community acquired infections. The K. pneumoniae infections have considerably contributed to morbidity and mortality in patients with protracted ailments. The capacity of K. pneumoniae to cause diseases depends on the presence of an array virulence factors. Coexistence and expression of virulence factors and genetic determinants of antibiotic resistance complicates treatment outcomes. Thus, emergence of pathogenic MDR K. pneumoniae poses a great threat to the healthcare system. However, the carriage of antibiotic resistance among pathogenic K. pneumoniae is yet to be investigated in Uganda. We sought to investigate the carbapenem resistance profiles and pathogenic potential based on capsular serotypes of K. pneumoniae clinical isolates. Methods This was a cross sectional study involving use of archived Klebsiella pneumoniae isolates collected between January and December, 2019 at four tertiary hospitals in Uganda. All isolates were subject to antimicrobial susceptibility assays to determine phenotypic antibiotic resistance, pentaplex PCR to detect carbapenemases encoding genes and heptaplex PCR to identify capsular serotypes K1, K2, K3, K5, K20, K54 and K57. Results The study found an overall phenotypic carbapenem resistance of 23.3% (53/227) and significantly higher genotypic resistance prevalence of 43.1% (98/227). Over all, the most prevalent gene was blaOXA-48-like (36.4%), followed by blaIMP-type (19.4%), blaVIM-type (17.1%), blaKPC-type (14.0%) and blaNDM-type (13.2%). blaVIM-type and blaOXA-48-like conferred phenotypic resistance in all isolates and 38.3% of isolates that harbored them respectively. Capsular multiplex PCR revealed that 46.7% (106/227) isolates were pathogenic and the predominantly prevalent pathotype was K5 (18.5%) followed by K20 (15.1%), K3 (7.1%), K2 (3.1%) and K1 (2.2%). Of the 106 capsular serotypes, 37 expressed phenotypic resistance; thus, 37 of the 53 carbapenem resistant K. pneumoniae were pathogenic. Conclusion The high prevalence of virulent and antibiotic resistant K. pneumoniae among clinical isolates obtained from the four tertiary hospital as revealed by this study pose a great threat to healthcare. Our findings underline the epidemiological and public health risks and implications of this pathogen.

Molecular Detection Of Some Capsulargenes Of Klebsiella Pneumoniae Isolated From Clinical Samples

“…Seven individual PCRs were performed to detect the presence of Capsule serotype genes( k1,k2,k3,k5,k20,k54,k57) in K. pneumonia isolates as in previous studies [1,20].Primer sequences, and amplicon sizes are shown in Table1.Amplified PCR products were analyzed by gel electrophoresis on 1.5% agarose gels containing Safe Red(0.005% v/v) in 1 × TAE buffer, and imaged using Gel documentation system(Wisd-Korea(. (3) .In the study of Ssekatawa et al [18], a variety of clinical specimens such as wound, burn, sputum and urine were used to study the prevalence of K. pneumonia in a hospital based study and they were able to isolate the microorganisms from all these samples supporting our finding that K. pneumonia has the ability to cause a wide spectrum of clinical illness and can be isolated from a variety of clinical samples. The samples of K. pneumonia obtained in the current study were taken from four main clinical patterns: burn, urine ,wound and sputum as shown in figure (1).…”

“…Regarding serotype K2 gene, it was identified in 12.9 % isolates of our study but was not detected in study of Hamam et al [10] and Shakib et al [14] . However, it was reported in 6.7 % in the study of Chen et al [5] Serotype K54 detected in 37.8 % isolates of our study and 2.1 % in the isolates of study by Chen et al [5] .Capsule serotype k57 in 25.8 % isolates of our study, while in the study of Ssekatawa et al [18] there was no detection of K57 serotype gene. The K genes are capsular virulence factors involved in highly virulent K. pneumonia [5] .…”

Molecular characterization and distribution of cephalosporin resistance determinants in Escherichia coli and Klebsiella pneumoniae isolated from patients attending Kampala International University Teaching Hospital in Bushenyi, Western Uganda

“…This resistance may be attributed to the presence of resistance intrinsic factors, such as downregulation and mutation of the outer membrane proteins, through which antibiotics diffuse [41][42][43][44]. High prevalence of carbapenem resistance has been reported in Uganda [45][46][47]. Acquisition of carbapenemase encoding genes by Enterobacteriaceae confer resistance to all beta lactam antibiotics [41,45].…”

Phyto-Mediated Copper Oxide Nanoparticles for Antibacterial, Antioxidant and Photocatalytic Performances

“…Furthermore, resistance to last-resort antibiotics used in treating multidrug-resistant (MDR) infections has been reported worldwide ( Centre for Disease Dynami, 2015 ). For example, several studies have reported a high prevalence of carbapenem resistance in Uganda ( Ssekatawa et al, 2018 ; Ssekatawa et al, 2021a ; Ssekatawa et al, 2021b ), yet carbapenems are a class of antibiotics earmarked to treat MDR gram-negative bacterial infections. Thus, resistance to carbapenems limits the treatment options for MDR infections.…”