Molecular Detection of Carbapenem Resistance Genes of Klebsiella pneumoniae Isolated from Clinical Specimens in Khartoum State

Abstract

Carbapenems are the most powerful β-lactam antibiotics against bacteria especially multidrug resistant isolates like Klebsiella pneumoniae (K. pneumoniae). Since there were no comprehensive studies about prevalence of carbapenem resistance genes in Sudan, so this study was conducted to detect carbapenem resistance genes of K. pneumoniae from clinical specimens in Khartoum State during the period from March to November 2018. A total of sixty (n=60) K. pneumoniae were included in this descriptive-cross sectional laboratory based study. K. pneumoniae were isolated from urine, blood, wound swab and sputum samples, which were obtained from different hospitals in Khartoum State. The samples were collected from both males and females with different age using non-self-constructing information list. The isolated organism was stored in 20% Glycerol peptone media and was inoculated in MacConkey’s agar. Antibiotic susceptibility tests were carried out using Kirby Bauer technique. DNA was extracted using guanidine chloride method. The isolates were detected using PCR based method targeting 16S rRNA gene then were tested for the presence of carbapenem resistance genes (NDM, IMP, SPM, OXA-48 and KPC) using Multiplex PCR. Out of 60 K. pneumoniae isolates, 48 (80%) were positive for one or more carbapenem resistance genes; 41 (68.3%) were positive for OXA-48 gene, 6 (10%) were positive for NDM gene, 5 (8.4%) were positive for SPM gene, 5 (8.4%) were positive for KPC gene and 2 (3.3%) were positive for IMP gene. Ten isolates possess more than one gene. Thirty-eight isolates possess one gene and the remaining twelve isolates were free from these genes. It is concluded that, there is a high percentage and multiplicity of carbapenem resistance genes among K. pneumoniae isolates in Khartoum State. In addition, this study shown for the first time, K. pneumoniae carried SPM gene.