{"id":66384,"date":"2025-06-30T10:10:29","date_gmt":"2025-06-30T10:10:29","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=66384"},"modified":"2025-07-16T05:58:04","modified_gmt":"2025-07-16T05:58:04","slug":"antimicrobial-susceptibility-and-molecular-detection-of-carbapenem-resistant-genes-ndm-1-and-imp-1-of-pseudomonas-aeruginosa-isolated-from-clinical-samples-in-khartoum-state-sudan","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol18no2\/antimicrobial-susceptibility-and-molecular-detection-of-carbapenem-resistant-genes-ndm-1-and-imp-1-of-pseudomonas-aeruginosa-isolated-from-clinical-samples-in-khartoum-state-sudan\/","title":{"rendered":"Antimicrobial Susceptibility and Molecular Detection of Carbapenem Resistant Genes NDM-1 and IMP-1 of Pseudomonas aeruginosa Isolated from Clinical Samples in Khartoum State-Sudan"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

P. eruginosa<\/em>, one of the most common opportunistic pathogen associated with nosocomial infections, including pneumonia, urinary tract infections, and wound infections. Although carbapenems are frequently used as a therapeutic agent for treating infections caused by P. aeruginosa<\/em>, the high emergence of carbapenem resistance significantly decreases their utility.1<\/sup> Resistance of Pseudomonas aeruginosa<\/em> to different classes of antibiotics such as penicillin, cephalosporin, quinolone, aminoglycoside and carbapenem. Metallo beta lactamases (MBL) are enzymes that catalyze the hydrolysis of broad-spectrum beta lactam antibiotics including carbapenems. Resistant to carbapenems especially in Gram negative pathogens is global public health issue because of the spread of transferable carbapenemase encoding genes. The antibiotic resistant elements could be transferred to other Gram-negative bacterial strains and contribute towards spread of antimicrobial resistance rate making the treatment of infected cases complex. 2<\/sup> Carbapenems are the antibiotics which are utilized for treating multidrug-resistant P.aeruginosa <\/em>(MDRP) isolates. Carbapenem antibiotics used to be effective agents against MDRP when first presented. However, the growing frequence of carbapenem resistant P. aeruginosa <\/em>(CRPA) has turned into a severe health problem recently. These strains lead to high mortality rates in cases infected by P. aeruginosa <\/em>and there are also few effective drugs against them. Resistance to carbapenems can be related to the production of carbapenemase enzymes such as serine carbapenemases (containing KPC and GES enzymes), metallo-\u03b2-lactamases (MBLs) such as IMP, VIM, and NDM enzymes, and oxacillinases (such as OXA enzymes). blaIMP and blaVIM are the most constantly acquired MBLs. However, the recently emerged NDM type (New Delhi metallo-\u03b2 lactamases) is getting the most important carbapenemase. 3<\/sup><\/p>\n

Carbapenems have become the drug of choice for the treatment of severe nosocomial infections caused by Gram-negative bacilli. Carbapenem resistant Enterobacteriales (CRE) is a considerable health problem globally and are associated with increased mortality\u00a0 also \u00a0\u00a0There is a significant challenge in controlling the spread of carbapenemases, Surveys of the molecular epidemiology of carbapenemases revealed that the dissemination of carbapenemases, including NDM, VIM, IMP, OXA-48 and KPC producers, are rapid and widespread among healthcare facilities. Molecular techniques remain the gold standard for the precise identification of carbapenemase genes. 4<\/sup> Most of these techniques are based on PCR and may be followed by a sequencing step if a precise identification of the carbapenemase gene is needed (e.g. VIM type, KPC type, NDM type or OXA-48 type). 4<\/sup> They are either single or multiplex PCR techniques.4 <\/sup><\/p>\n

In Africa, data on the frequence and distribution of carbapenem resistance among the multidrug resistant Gram-negative bacteria is still limited.5<\/sup> In addition, detection discovery of carbapenem resistance genes producing bacteria may be difficult based on routine antibiotic susceptibility testing.6 <\/sup>Considering the fact that there were no comprehensive studies about prevalence of carbapenem resistance genes in Sudan so detection of those genes in P. aeruginosa<\/em> from clinical samples will be of a great value. Such data serve an important part in understanding the spread of this pathogen and help in good treatment for it. So this study aimed to detect carbapenem resistance genes of Pseudomonas aeruginosa<\/em> from clinical samples in Khartoum State, using conventional and PCR based method.<\/p>\n

Materials and Methods<\/strong><\/p>\n

Study sample and data collection<\/strong><\/p>\n

A total of fifty non-repetitive P.aeruginosa <\/em>(n=50),\u00a0 isolated from different clinical specimens<\/p>\n

Data included sample type, gender, and age of patients were collected from hospitals’ medical records checklist.<\/p>\n

Isolation and identification of bacteria <\/strong><\/p>\n

One colony from each isolate was sub-cultured on Cysteine Lactose Electrolyte Deficient media (CLED) and Nutrient agar and incubated aerobically at 37\u00b0C overnight, for further Re-identification and preservation. \u00a0P.aeruginosa<\/em>\u00a0showed\u00a0smooth blue non-lactose fermenting colonies on CLED, were identified by Gram stain and conventional biochemical tests.<\/p>\n

Antimicrobial Susceptibility test<\/strong><\/p>\n

All confirmed isolates were subculture from 15% glycerol on nutrient agar getting overnight and pure growth. Modified Kirby Bauer was performed and interpreted according to the Clinical and Laboratory Standards Institute (CLSI, 2022).\u00a0<\/strong><\/p>\n

DNA Extraction<\/strong><\/p>\n

The boiling method was used for isolation of bacterial genomic DNA which extracted from fresh overnight incubated isolate that suspended in 200ul of d H2O in 1.5 eppendrof tube, bolied for 10 min at 100\u00b0C in thermal block incubator, vortex, then cooling at -20\u00b0C following by centrifuged for 10 min at 12000xg. Supernatant were carefully collected and store at -20\u00b0C until analyzed. The purity of the extracted DNA was determined by running the DNA sample on 1.5% agarose gel.<\/p>\n

Polymerase Chain Reaction Amplification<\/strong><\/p>\n

PCR was done by multiplex PCR, amplification which done using Maxime PCR Premix kit and specific primer which sensitized by Specific primers were used to amplification indicated in table (1). The PCR assay was carried out in a total volume of 25 \u03bcL of mixture containing 2 \u03bcLMaxime PCR Premix containing 1X PCR buffer, 1.5 mM MgCl2, 200M \u03bc of each dNTP, and 1 U Taq DNA polymerase, 0.5 \u03bcL of each of the virulence gene-specific primers (2 \u03bcL), 5 \u03bcL of template DNA and 13 \u03bcL of water for injection. The amplification conditions included three steps: heating at 95\u00b0C for 5 min; 34 cycles of denaturation at 95\u00b0C for 1 min, annealing at 58\u00b0C for 1min for (NDM -1 and IMP -1 genes) and extension at 72\u00b0C for 1min; and the final extension at 72\u00b0C for 5 min. 7<\/sup><\/p>\n

Table 1: Primers used for amplification of carbapenem resistance genes of P.aeruginosa<\/em> isolates <\/strong><\/p>\n\n\n\n\n\n\n\n
Target gene<\/strong><\/td>\nSequence (5′- 3′)<\/strong><\/td>\nAmplicon size (bp)<\/strong><\/td>\n<\/tr>\n
IMP-F<\/strong><\/td>\nGGAATAGAGTGGCTTAAYTC<\/td>\n\u00a0<\/strong>\u00a0<\/strong><\/p>\n

Bla IMP -1 232<\/strong><\/td>\n<\/tr>\n

IMP-R<\/strong><\/td>\nGGTTTAAYAAAACAACCACC<\/td>\n<\/tr>\n
NDM-F<\/strong><\/td>\nGGTTTGGCGATCTGGTTTTC<\/td>\n\u00a0<\/strong>\u00a0<\/strong><\/p>\n

Bla NDM -1621<\/strong><\/td>\n<\/tr>\n

NDM-R<\/strong><\/td>\nCGGAATGGCTCATCACGATC<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Gel electrophoresis <\/strong><\/p>\n

The gel casting tray was put into the electrophoresis, tank flooded with 1x TBE buffer just to cover the gel surface, 2\u03bcl of PCR products from each 50samples was added to wells of electrophoreses, 5 \u03bcl of100-bp DNA ladder, was added to the well in each run. The electrophoresis was carried out at 120Volts for 2 Hours. Then the gel was visualized for DNA bands by U.V transilluminator and photographed.<\/p>\n

Statistical analysis<\/strong><\/p>\n

All outcome data were analyzed by chi-square using Statistical Package for Social Sciences (SPSS; Version16).<\/p>\n

Results<\/strong><\/p>\n

Out of the 50 isolates, 18 were isolated from urine samples, 11 from wound swabs, 6 from blood, 2 from high vaginal swabs, 8 from sputum, 3 from ear swabs, and 2 from body fluids, after culturing most strains were show pigmentation 32 (74%), the most pigmented strain was found in urine (26%) and wound samples (20%) table (2). Demographic data show out of 50 the male 23 were (46%),where female were 27 (54%) with ratio1 : 0.85. Ages ranged from 4 to 76 years, with an average of 49. Majority of the samples (37%) were found in the age group (40 \u2013 60) year table (3).<\/p>\n

Approximately one-third 18 (36%) of isolates were resistant to Aztreonam, 10 (20%) were resistant to Ciprofloxacine and Colistin, 11 (22%) resistant to Gentamicin, Amikacin, and Pipracillin\/tazobactam combinations, only one isolate was resistant to Imipenem, from 18 urine samples 3 (19%) isolates were resistant to Norfloxacin table (4).<\/p>\n

Out of 50 P. aeruginosa<\/em> isolates; only 14 (28%) were MDR according to the preliminarily described definition , most isolates were sensitive to all antibiotics 10 (36%), 12 (24%) isolates resist to one antibiotic, 6 (12%) isolates resist to two antibiotics. In MDR isolates we found two strains were resistant to >7 antibiotics, whereas just three isolates were found to be resistant to 6 antibiotics. There was insignificant Association between sex and antimicrobial susceptibility results table (5). The NDM-1gene was detected in 6 isolates (12%) and negative in 44 isolate (88%). They were recovered from urine (8%), wound (4%), ear swab (0%), sputum (0%), blood (0%) high vaginal swabs (0%) and body fluid (0%) table (6). While the IMP-1 gene was detected in 13(26%), and negative in 37(74%) isolate table (6). There was insignificant difference in the percentage of resistant genes among the different infection site.\u00a0 The association between the presence of NDM-1 and IMP-1genes and antibiotics was significant with Pipracillin, Impenem, Aztreonam, Ceftazidime table (7).<\/p>\n

Table 2: Frequency of pigmented strains among different clinical samples<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/td>\nPigment appearance<\/strong><\/td>\n<\/tr>\n
Green Pigment<\/strong><\/td>\nNon pigmented<\/strong><\/td>\n<\/tr>\n
\u00a0<\/strong>\u00a0<\/strong><\/p>\n

\u00a0<\/strong><\/p>\n

Sample<\/strong><\/td>\n

Urine sample<\/td>\n13 (26%)<\/td>\n5 (10%)<\/td>\n<\/tr>\n
Sputum<\/td>\n7 (14%)<\/td>\n1 (2%)<\/td>\n<\/tr>\n
Wound<\/td>\n10 (20%)<\/td>\n1 (2%)<\/td>\n<\/tr>\n
Blood<\/td>\n4 (8%)<\/td>\n2 (4%)<\/td>\n<\/tr>\n
Ear swab<\/td>\n1 (2%)<\/td>\n2 (4%)<\/td>\n<\/tr>\n
High vaginal swab<\/td>\n2 (4%)<\/td>\n0 (0%)<\/td>\n<\/tr>\n
Body fluids<\/td>\n1 (2%)<\/td>\n1 2%)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table 3: Distribution of positive samples among different gender and age groups<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/td>\nGender<\/strong><\/td>\n\u00a0<\/strong><\/td>\n<\/tr>\n
Male<\/strong><\/td>\nFemale<\/strong><\/td>\nTotal<\/strong><\/td>\n<\/tr>\n
\u00a0<\/strong>Age group<\/strong><\/td>\n<20<\/td>\n1 (2%)<\/td>\n0 (0%)<\/td>\n1(2%)<\/td>\n<\/tr>\n
20-40<\/td>\n3 (6%)<\/td>\n10 (20%)<\/td>\n13 (26%)<\/td>\n<\/tr>\n
40-60<\/td>\n10 (20%)<\/td>\n10 (20%)<\/td>\n20 (40%)<\/td>\n<\/tr>\n
>60<\/td>\n9 (18%)<\/td>\n7 (14%)<\/td>\n16 (32%)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong>Table 4 : Antimicrobial susceptibility pattern of P . aeruginosa <\/em>isolates<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Sample\u00a0 <\/strong><\/td>\nUrine<\/strong>(N=18)<\/strong><\/td>\nWound swab<\/strong>(n=11)<\/strong><\/td>\nBlood<\/strong>(N=6)<\/strong><\/td>\nHVS<\/strong>(N=2)<\/strong><\/td>\nSputum<\/strong>(N=8)<\/strong><\/td>\nEar swab<\/strong>(N=3)<\/strong><\/td>\nBody fluid<\/strong>(N=2)<\/strong><\/td>\nTotal<\/strong>(N=50)<\/strong><\/td>\n<\/tr>\n
\u00a0<\/strong>Antibiotic<\/strong><\/td>\n<\/tr>\n
CIP<\/td>\nResistant<\/td>\n3<\/td>\n3<\/td>\n2<\/td>\n0<\/td>\n1<\/td>\n2<\/td>\n0<\/td>\n11(22%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n15<\/td>\n8<\/td>\n4<\/td>\n2<\/td>\n7<\/td>\n1<\/td>\n2<\/td>\n39(78%)<\/td>\n<\/tr>\n
CN<\/td>\nResistant<\/td>\n4<\/td>\n5<\/td>\n2<\/td>\n0<\/td>\n2<\/td>\n1<\/td>\n1<\/td>\n15(30%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n14<\/td>\n6<\/td>\n4<\/td>\n2<\/td>\n6<\/td>\n2<\/td>\n1<\/td>\n35(70%)<\/td>\n<\/tr>\n
TPZ<\/td>\nResistant<\/td>\n5<\/td>\n3<\/td>\n0<\/td>\n0<\/td>\n1<\/td>\n2<\/td>\n0<\/td>\n11(22%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n13<\/td>\n8<\/td>\n6<\/td>\n2<\/td>\n7<\/td>\n1<\/td>\n2<\/td>\n39(78%)<\/td>\n<\/tr>\n
CAZ<\/td>\nResistant<\/td>\n6<\/td>\n4<\/td>\n3<\/td>\n0<\/td>\n2<\/td>\n1<\/td>\n1<\/td>\n17(34%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n12<\/td>\n7<\/td>\n3<\/td>\n2<\/td>\n6<\/td>\n2<\/td>\n1<\/td>\n33(66%)<\/td>\n<\/tr>\n
ATM<\/td>\nResistant<\/td>\n12<\/td>\n4<\/td>\n3<\/td>\n1<\/td>\n7<\/td>\n1<\/td>\n1<\/td>\n30(60%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n6<\/td>\n7<\/td>\n3<\/td>\n1<\/td>\n1<\/td>\n2<\/td>\n1<\/td>\n20(40%)<\/td>\n<\/tr>\n
AK<\/td>\nResistant<\/td>\n2<\/td>\n5<\/td>\n1<\/td>\n0<\/td>\n3<\/td>\n1<\/td>\n1<\/td>\n13(26%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n16<\/td>\n6<\/td>\n5<\/td>\n2<\/td>\n5<\/td>\n2<\/td>\n1<\/td>\n37(74%)<\/td>\n<\/tr>\n
IMP<\/td>\nResistant<\/td>\n2<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n2 (4%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n16<\/td>\n11<\/td>\n6<\/td>\n2<\/td>\n8<\/td>\n3<\/td>\n2<\/td>\n48(96%)<\/td>\n<\/tr>\n
CT<\/td>\nResistant<\/td>\n5<\/td>\n1<\/td>\n0<\/td>\n1<\/td>\n2<\/td>\n1<\/td>\n0<\/td>\n10(20%)<\/td>\n<\/tr>\n
Sensitive<\/td>\n13<\/td>\n10<\/td>\n6<\/td>\n1<\/td>\n6<\/td>\n2<\/td>\n2<\/td>\n40(80%)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong>Table 5: Association between sex and Antimicrobial susceptibility results<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
\u00a0<\/strong>Antibiotic<\/strong><\/td>\nSex group<\/strong><\/td>\n<\/tr>\n
male<\/strong><\/td>\nFemale<\/strong><\/td>\n<\/tr>\n
Sensitive<\/strong><\/td>\nResistant<\/strong><\/td>\nSensitive<\/strong><\/td>\nResistant<\/strong><\/td>\n<\/tr>\n
CAZ<\/td>\n15<\/td>\n8<\/td>\n18<\/td>\n9<\/td>\n<\/tr>\n
AZT<\/td>\n11<\/td>\n12<\/td>\n10<\/td>\n17<\/td>\n<\/tr>\n
TPZ<\/td>\n17<\/td>\n6<\/td>\n22<\/td>\n5<\/td>\n<\/tr>\n
CT<\/td>\n21<\/td>\n2<\/td>\n19<\/td>\n8<\/td>\n<\/tr>\n
CN<\/td>\n13<\/td>\n10<\/td>\n22<\/td>\n5<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0Table 6: Association between P . aeruginosa <\/em>Resistant genes and infection sites<\/strong><\/p>\n\n\n\n\n\n\n
Resistant genes<\/strong><\/td>\nSample sites<\/strong><\/td>\n<\/tr>\n
Urine<\/strong>N=18<\/strong><\/td>\nWound<\/strong>N=11<\/strong><\/td>\nEar swab<\/strong>N=3<\/strong><\/td>\nBlood<\/strong>N=6<\/strong><\/td>\nBody fluid<\/strong>N=2<\/strong><\/td>\nSputum<\/strong>N=8<\/strong><\/td>\n<\/tr>\n
NDM-1<\/strong><\/td>\n4<\/td>\n2<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n0<\/td>\n<\/tr>\n
IMP-1<\/strong><\/td>\n6<\/td>\n4<\/td>\n0<\/td>\n1<\/td>\n1<\/td>\n1<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong>Table 7: Association between the presence of IMP-1gene and NDM -1 gene WITH antibiotics susceptibility<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Antibiotic<\/strong><\/td>\nIMP -1<\/strong><\/td>\nNDM -1<\/strong><\/td>\n<\/tr>\n
(+ve)<\/strong><\/td>\n(-ve)<\/strong><\/td>\n(+ve)<\/strong><\/td>\n(-ve)<\/strong><\/td>\n<\/tr>\n
Ciprofloxacin<\/td>\nR<\/td>\n5<\/td>\n12<\/td>\n2<\/td>\n15<\/td>\n<\/tr>\n
S<\/td>\n8<\/td>\n25<\/td>\n4<\/td>\n29<\/td>\n<\/tr>\n
Colistin<\/td>\nR<\/td>\n3<\/td>\n10<\/td>\n3<\/td>\n10<\/td>\n<\/tr>\n
S<\/td>\n10<\/td>\n27<\/td>\n3<\/td>\n34<\/td>\n<\/tr>\n
Gentamicin<\/td>\nR<\/td>\n4<\/td>\n18<\/td>\n2<\/td>\n20<\/td>\n<\/tr>\n
S<\/td>\n9<\/td>\n19<\/td>\n4<\/td>\n24<\/td>\n<\/tr>\n
Amikacin<\/td>\nR<\/td>\n1<\/td>\n5<\/td>\n0<\/td>\n6<\/td>\n<\/tr>\n
S<\/td>\n12<\/td>\n32<\/td>\n6<\/td>\n38<\/td>\n<\/tr>\n
Pipracillin<\/td>\nR<\/td>\n7<\/td>\n19<\/td>\n1<\/td>\n25<\/td>\n<\/tr>\n
S<\/td>\n6<\/td>\n18<\/td>\n5<\/td>\n19<\/td>\n<\/tr>\n
Impenem<\/td>\nR<\/td>\n5<\/td>\n18<\/td>\n2<\/td>\n21<\/td>\n<\/tr>\n
S<\/td>\n8<\/td>\n19<\/td>\n4<\/td>\n23<\/td>\n<\/tr>\n
Aztreonam<\/td>\nR<\/td>\n8<\/td>\n24<\/td>\n3<\/td>\n29<\/td>\n<\/tr>\n
S<\/td>\n5<\/td>\n13<\/td>\n3<\/td>\n15<\/td>\n<\/tr>\n
Ceftazidime<\/td>\nR<\/td>\n10<\/td>\n26<\/td>\n3<\/td>\n33<\/td>\n<\/tr>\n
S<\/td>\n3<\/td>\n11<\/td>\n3<\/td>\n11<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\nFigure 1: Gel electrophoresis of NDM -1 gene. The gene was detected in 6 isolates out of the 50 isolates. Lane: 9 and 18 ladder 100 bp. Lane: 1\/5\/12\/13\/15\/and 17 positives for IMP -1 product (232bp). Lane:7\/8 and 11 show negative results. Lane: 1\/2\/4\/13 \/14 and16 show positive for NDM -1product (621bp).<\/span><\/strong>Click here to view Figure\u00a0<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\nFigure 2: Gel electrophoresis of IMP -1 gene, the gene was harbored in13samples Lane: 9 ladder 100 bp. Lane: 1\/4\/5and 6 positive for NDM -1 product (621bp). Lane: 2\/7 show negative results. Lane: 1\/3and 8 positive for IMP -1 product\u00a0\u00a0 (232bp).<\/strong>Click here to view Figure\u00a0<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Discussion <\/strong><\/p>\n

In the current study, most of isolates harboring carbapenem resistance genes were phenotypically susceptible to imipenem and most of them were positive for IMP gene. This confirm what 8<\/sup> said that this gene is not strong and relies upon other synergistic mechanisms to mediate resistance against carbapenems. In addition to imipenem other antibiotics were analyzed in this study. Majority of the isolates showed varying degrees of resistance to the other antibiotics (ciprofloxacin, gentamicin and ceftazidime). Resistance to these antibiotics may also be due to the presence of ESBLs and other mechanisms like efflux pumps and porin mutations 9<\/sup>, which were not covered in this study.<\/p>\n

In the current study, the most prevalent gene among the\u00a0 50P. aeruginosa<\/em> isolates was IMP\u00a0 gene (26 %). NDM gene was\u00a0 also present but on a smaller scale (12%, n =6 ). This is consistent with previous findings by in Turkey10<\/sup>, which detected OXA-48 gene (58%) and NDM gene (2%) 11<\/sup>,\u00a0 detected OXA-48 gene (64.7%) and NDM gene (5.8%) and 12 <\/sup>detected OXA-48 gene (78%) and NDM gene (20%) both from Saudi Arabia, IMP gene were not detected in these studies<\/p>\n

These variations in results may be because, they only detect resistant genes in carbapenem-resistant isolates or due to differences in method of detection as the first one used real-time PCR or differences in the sample size of tested organism and that was seen with the other two studies.<\/p>\n

Of 50 P. aeruginosa <\/em>isolates detected of having carbapenem resistance genes, two had multiple genes. This finding agree with 13<\/sup> and 6 <\/sup>which showed multiplicity of genes in their isolates. The presence of multiple resistance genes in one isolate have not been generally detected in a large number of studies probably due to the limited number of genes studied since most of the studies research on one or two genes. 6<\/sup> Regarding NDM and IMP gene, 6 (12%) and 13(26%) were positive respectively. Ceftazidime resistant Isolates showed significant association with both NDM and IMP gene as detected 3, 10 isolates respectively. One of the limitations of this study; it was conducted on a limited number of samples so future studies should be carried with large sample size and different clinical isolates to and sequencing technique to document the prevalence of these resistant genes and to help on therapeutic strategies.<\/p>\n

Conclusion<\/strong><\/p>\n

In conclusion, the findings of the present study disclosed a high percentage andmultiplicity of carbapenem resistance genes among P. aeruginosa <\/em>isolates in Khartoum State. This study showed that the NDM-1 and IMP-1 genes were generally circulated among the P. aeruginosa.<\/em><\/p>\n

Acknowledgment<\/strong><\/p>\n

The authors would like to thank all the colleagues in Microbiology Laboratory, College of Medical Laboratory, Sudan University of sciences and technology who helped in the study, and also our thanks to Dr. Osama Mohamed a leader of EXON lab of molecular biology for his guidance and help in molecular work.<\/p>\n

Funding Sources<\/strong><\/p>\n

The author(s) received no financial support for the research, authorship, and\/or publication of this article.<\/p>\n

Conflect of Interest<\/strong><\/p>\n

The author(s) do not have any conflict of interest<\/p>\n

Data Availability<\/strong><\/p>\n

This statement does not apply to this article.<\/p>\n

Ethics Statement<\/strong><\/p>\n

This research did not involve human participants, animal subjects, or any material that requires ethical approval<\/p>\n

Informed Consent Statement<\/strong><\/p>\n

This study did not involve human participants, and therefore, informed consent was not required<\/p>\n

Clincal Trial Registeration<\/strong><\/p>\n

This research does not involve any clinical trials.<\/p>\n

Permission to Reproduce Material from Other Source<\/strong><\/p>\n

Not Applicable<\/p>\n

Authors Contributions\u00a0 <\/strong><\/p>\n