Incidence of Drug-Resistant Hospital-Associated Gram-Negative Bacterial Infections, the Accompanying Risk Factors, and Clinical Outcomes with Treatment.

Extensive drug resistance to bacterial infections in hospitalised patients is accompanied by high morbidity and mortality rates due to limited treatment options. This study investigated the clinical outcomes of single and combined antibiotic therapies in extensive (XDR), multidrug-resistant (MDR) and susceptible strains (SS) of hospital-acquired infections (HAIs). Cases of hospital-associated drug-resistant infections (HADRIs) and a few susceptible strains from hospital wards were selected for this study. Bacteria identifications (IDs) and antimicrobial susceptibility tests (ASTs) were performed with a Vitek 2 Compact Automated System. Patients' treatment types and clinical outcomes were classified as alive improved (AI), alive not improved (ANI), or died. The length of hospital stay (LOHS) was acquired from hospital records. The HAI pathogens were Acinetobacter baumannii (28%), Escherichia coli (26%), Klebsiella pneumoniae (22%), Klebsiella (2%) species, Pseudomonas aeruginosa (12%), Proteus mirabilis (4%), and other Enterobacteriaceae. They were MDR (40.59%), XDR (24.75%), carbapenem-resistant Enterobacteriaceae (CRE, 21.78%) and susceptible (12%) strains. The treatments were either monotherapy or combined therapy with different outcomes. Monotherapy produced positive significant outcomes with E. coli infections, while for P. aeruginosa, there were no differences between the number of infections treated with either mono/combined therapies (50% each). Nonetheless, combined therapy had significant effects (p < 0.05) as a treatment for A. baumannii and K. pneumoniae infections. Clinical outcomes and LOHS varied with infecting bacteria. The prevalence of XDR and MDR HAIs was found to be significantly high, with no association with treatment type, LOHS, or outcome.

Diagnosis of group A streptococcal pharyngitis in the paediatric emergency department using a fluorescence-based RADT: predicted impact on antibiotic prescription.

Introduction. Using rapid antigen diagnostic tests (RADTs) in clinical practice has shown excellent specificity but often has diminished sensitivity.Gap Statement. Local data for evaluating the diagnostic performance of a new fluorescence-based RADT and its influence on the antibiotic prescription rate are not available.Aim. To evaluate the accuracy of fluorescent immunoassay (FIA)-RADTs for diagnosing group A streptococcal (GAS) pharyngitis among children and its estimated effect as a point of care test (POCT) on the antibiotic prescription rate at the paediatric emergency department.Methodology. A prospective study was conducted, comprising children 3 to 14 years old presenting with pharyngitis. Throat swab culture and FIA-RADTs were performed on all samples. Conventional PCR was performed on the discordant samples.Results. A total of 246 children were included in this study. The sensitivity, specificity, and positive and negative predictive values of the FIA-RADT, based on culture results and PCR detection combined, were 95.6, 96.8, 94.6 and 97.4 %, respectively. Antibiotics have been prescribed to 162 (65.9 %) children; however, if FIA-RADTs had been added in the clinical practice as a POCT, only 92 (37.4 %) children would have received antibiotics in total. Additionally, implementation of FIA-RADTs would significantly reduce the antibiotic prescription rate from 48.8 and 60.6 % to 9.5 and 31.9 % among patients with clinical scores of 2 and 3, respectively.Conclusion. The new FIA-RADT is simple, prompt and reliable. It is helpful in clinical settings and may be used to reduce antibiotic overprescription, especially for children who have a low risk for GAS pharyngitis, according to the clinical score.

Molecular characterization of Helicobacter pylori clinical isolates from Eastern Saudi Arabia.

OBJECTIVES: To describe the frequency of cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) virulence genes and clarithromycin resistance-associated mutations among Helicobacter pylori (H. pylori) clinical isolates from Eastern Saudi Arabia. METHODS: A cross-sectional study was carried out between July 2020 and June 2021 in a tertiary hospital in AL-Khobar, Saudi Arabia. A total of 34 H. pylori isolates were obtained from gastric biopsies of patients with dyspepsia. The existence of the virulence genes was studied by polymerase chain reaction and the gene fragment of the 23s ribosomal subunit (23s rRNA) gene was sequenced. RESULTS: All isolates harbored the CagA gene. Approximately 97.1% (33/34) isolates were positive using the VacA M primer and 91.2% (31/34) isolates were positive using the VacA S primer. The most frequent allelic combination was S2/M2/cag (60%), followed by S1/M2/cag (26.7%), S1/M1/cag (10%), and S2/M1/cag (3.3%). Approximately 6.5% isolates harbored the A2142G mutation and 29% isolates harbored the A2143G mutation. One isolate contained the mutation T2182C. The phylogenetic analysis showed that 58% isolates clustered with the regional and global isolates while the remaining 42% isolates seemed to be specifically circulating in Saudi Arabia. Most of the patients (73.5%) had already underwent a previous H. pylori eradication therapy. CONCLUSION: We showed that there is a regional variation in the frequency of the virulence genes among H. pylori isolates. Additionally, we showed the frequency of 23s rRNA mutations related to clarithromycin resistance in Saudi Arabia.

Low levels of soluble DPP4 among Saudis may have constituted a risk factor for MERS endemicity.

Most of the cases of Middle East respiratory syndrome coronavirus (MERS-CoV) were reported in Saudi Arabia. Dipeptidyl peptidase-4 (DPP4) was identified as the receptor for the virus. The level of soluble DPP4 (sDPP4) was found to be reduced in MERS-CoV infected patients while high levels of sDPP4 were suggested to be protective against MERS-CoV in animal models. We investigated whether the Saudi population has lower levels of sDPP4 which makes them more susceptible to MERS-CoV infection and, therefore, could explain the larger number of cases from the country. Blood samples were collected from 219 Saudi blood donors and 200 blood donors from other ethnic groups. The plasma level of sDPP4 was measured by ELISA and the following SNPs in the DPP4 gene; rs35128070, rs1861978, rs79700168, and rs17574, were genotyped by TaqMan SNP genotyping assay. The average level of plasma sDDP4 was significantly lower in Saudis than other Arabs and non-Arabs (P value 0.0003 and 0.012, respectively). The genotypes AG of rs35128070 and GT of rs1861978 were significantly associated with lower sDPP4 among Saudis (P value 0.002 for each). While both genotypes AA and AG of rs79700168 and rs17574 were associated with significantly lower average sDPP4 level in Saudis compared to other ethnic groups (P value 0.031 and 0.032, and 0.027 and 0.014, respectively). Herein, we report that the Saudi population has lower levels of plasma sDPP4 than other ethnic groups, which is associated with genetic variants in the DPP4 gene. This may have contributed to increase the susceptibility of the Saudi population to MERS-CoV infection and could be a factor in the long-lasting persistence of the virus in the country.

Investigation of nonsynonymous mutations in the spike protein of SARS-CoV-2 and its interaction with the ACE2 receptor by molecular docking and MM/GBSA approach.

COVID-19 is an infectious and pathogenic viral disease caused by SARS-CoV-2 that leads to septic shock, coagulation dysfunction, and acute respiratory distress syndrome. The spreading rate of SARS-CoV-2 is higher than MERS-CoV and SARS-CoV. The receptor-binding domain (RBD) of the Spike-protein (S-protein) interacts with the human cells through the host angiotensin-converting enzyme 2 (ACE2) receptor. However, the molecular mechanism of pathological mutations of S-protein is still unclear. In this perspective, we investigated the impact of mutations in the S-protein and their interaction with the ACE2 receptor for SAR-CoV-2 viral infection. We examined the stability of pathological nonsynonymous mutations in the S-protein, and the binding behavior of the ACE2 receptor with the S-protein upon nonsynonymous mutations using the molecular docking and MM_GBSA approaches. Using the extensive bioinformatics pipeline, we screened the destabilizing (L8V, L8W, L18F, Y145H, M153T, F157S, G476S, L611F, A879S, C1247F, and C1254F) and stabilizing (H49Y, S50L, N501Y, D614G, A845V, and P1143L) nonsynonymous mutations in the S-protein. The docking and binding free energy (ddG) scores revealed that the stabilizing nonsynonymous mutations show increased interaction between the S-protein and the ACE2 receptor compared to native and destabilizing S-proteins and that they may have been responsible for the virulent high level. Further, the molecular dynamics simulation (MDS) approach reveals the structural transition of mutants (N501Y and D614G) S-protein. These insights might help researchers to understand the pathological mechanisms of the S-protein and provide clues regarding mutations in viral infection and disease propagation. Further, it helps researchers to develop an efficient treatment approach against this SARS-CoV-2 pandemic.

mCIM test as a reliable assay for the detection of CRE in the Gulf region.

Introduction. Carbapenem resistant Enterobacterales (CRE) are one of the leading causes of systemic and nosocomial infections and are multidrug-resistant organisms producing different carbapenemases. There are many genotypic and phenotypic methods for detecting the carbapenemases; however, there is a limitation for each. Modified carbapenem inactivation method (mCIM) assay is a recent phenotypic method which has been published by the Clinical and Laboratory Standards Institute.Hypothesis / Gap Statement. mCIM assay could provide a reliable method for the detection of carbapenemases in CRE.Aim. Evaluation of the mCIM assay performance for the detection of carbapenemases in Enterobacterales and the identification of the common carbapenemase genes at Eastern Province of Saudi Arabia and Kingdom of Bahrain.Methodology. A collection of 197 non-duplicate carbapenem resistant Enterobacterales clinical isolates, were evaluated with the mCIM test comparing its performance to multiplex PCR. The minimum inhibitory concentration susceptibility testing was done by the Etest method for imipenem, meropenem, and ertapenem.Results. The sensitivity of the mCIM assay was 94 % (95 % CI, (89.3-97.1)). In Saudi Arabia and Bahrain, OXA-48 was the most prevalent carbapenemase gene followed by NDM. Coexistence of multiple carbapenemase genes is reported in eleven cases.Conclusion. These findings indicate that the mCIM test is a reliable and simple assay for detecting the activity of carbapenemase in Enterobacterales, especially in resource-limited laboratories.

Pooled Specimen Testing Using Automated Cartridge-Based System for COVID-19: The Cost on Sensitivity.

BACKGROUND: Pooled specimen screening for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can improve laboratory testing capacity. This study assessed the impact of pooling and retesting individual swabs on the overall detection rate and reduction in the frequency of retesting. METHODS: One hundred respiratory swabs specimens were tested individually and in pools of three or five samples using the Cepheid's Xpert® Xpress SARS-CoV-2 test kit. The optimum number of samples per pool was calculated using the application 'A Shiny App for Pooled Testing'. RESULTS: Twenty-five pools were generated from 101 samples. Out of 13 pools that contained five samples each, three pools gave true positive results. While out of the 12 pools that contained three samples each, five pools gave true positive results. Four samples gave a false negative pool result. The overall sensitivity and specificity of the assay in the pools were 66.6% and 100%, respectively. The cycle threshold was reduced in most of the pools compared to individual sample tests. CONCLUSION: The overall pooled test had a remarkable impact on laboratory resources. Yet, caution is warranted when selecting the cases for pooled testing, since the reduction in sensitivity can significantly impact and increase the risk of exposure to infection.

Surgical site infections in a tertiary hospital over 10 years. The effect of hospital accreditation strategy implementation.

OBJECTIVES:   To analyze the rate of surgical site infections (SSIs), the type, and the frequency of the commonly-associated microorganisms. Methods: A retrospective study was conducted in King Fahd Hospital of the University, Al Khobar, Saudi Arabia between December 2018 and June 2019 comprising data from August 2008 to August 2018 from patients with culture-confirmed surgical site infection at a tertiary hospital. Results: Out of 2716 wound infection cases during the study period, a total of 289 patients were diagnosed with SSI. The rate of SSI in the tertiary hospital decreased from 20 per 1000 operations in 2009 to 3.5 per 1000 operations in 2018. A significant decrease in the rate of SSIs was observed  in 2014 and 2015 when self-assessment strategies in preparation for the accreditation of the hospital were implemented. A significant shift in the SSI rate from type I and II wounds to type IV wounds was observed coinciding with implementation of accreditation procedures. Escherichia coli was the most common pathogen. Antibiotic susceptibility patterns showed reduced resistance to ceftazidime and tazocin, while Acinetobacter baumannii was resistant to most of the antibiotics over 10 years. CONCLUSION: This study describes, for the first time, the status of SSI over the past 10 years in Saudi Arabia. The study also demonstrated the effect of hospital accreditation on healthcare organization performance regarding infection control and antibiogram pattern.

Pattern of increased antimicrobial resistance of Salmonella isolates in the Eastern Province of KSA.

OBJECTIVES: The antimicrobial resistance of Salmonella species is increasing worldwide. This study was conducted to determine the pattern of antimicrobial susceptibility of Salmonella species in a tertiary hospital from 2011 to 2018. METHODS: In this retrospective study, the medical records of all patients with Salmonella infections were reviewed. The clinical, demographic, and microbiological data of the selected patients were analysed. RESULTS: A total of 752 patients were included. The resistance of Salmonella species to antimicrobial drugs increased from 24.6% in 2011 to 37.8% in 2018 (p = 0.002). By 2018 all Salmonella isolates were completely resistant to cefalotin, cefuroxime, and cefoxitin, while we found some susceptibility to other cephalosporins and ciprofloxacin. The most commonly isolated Salmonella serogroups were groups D (36.5%), C (23.5%), and B (11.7%). CONCLUSIONS: The incidence of resistance of Salmonella to antibiotics is on the rise. The results of this study highlight the need for an active monitoring system of antibiotic usage in humans and domestic animals.

Association between Hepatitis C Virus Viremia and the rs12979860, rs2228145 and rs1800795 SNP (CT/AC/GG) Genotype in Saudi Kidney Transplant Recipients.

BACKGROUND: Hepatitis C virus (HCV) is a major health problem, particularly in high-risk groups such as kidney transplant recipients, where it can adversely affect graft survival and increase the relative risk for mortality. Recently, the role of genetic variation among HCV patients in determining the outcome of infections has been under investigation. OBJECTIVE: To investigate the association of single-nucleotide polymorphisms (SNPs) rs12979860 (located within the interleukin-28B locus), rs2228145 (interleukin-6 receptor) and rs1800795 (interleukin-6 promoter) with HCV viremia in renal transplant patients. MATERIALS AND METHODS: In this analytical cross-sectional study, 149 kidney transplant recipients, 82 males (median age: 41 years) and 67 females (median age: 45 years), were screened for HCV RNA in blood using real-time polymerase chain reaction and genotyped by sequencing (rs12979860) and restriction fragment length polymorphism (rs2228145 and rs1800795). RESULTS: HCV RNA was detected in 17 (11.41%) of the 149 patients. There was no statistically significant association between the studied SNPs and HCV viremia. However, a combination of the CT/AC/GG genotype was significantly associated with HCV viremia (odds ratio: 5.4). The genotype AA of rs2228145 in the IL-6 receptor was associated with viremia levels of >105 copies/ml (odds ratio: 5.96). CONCLUSION: To the best of the authors' knowledge, this is the first study that has shown that the CT/AC/GG genotype has an impact on HCV viremia in kidney transplant patients. Therefore, such SNP genotypes may potentially be used to identify transplant patients at risk of HCV infection.

First description of plasmid mediated quinolone resistance genes in salmonella isolates from Saudi hospitals.

OBJECTIVES: To detect resistance genes to fluoroquinolones and ß-lactams in Salmonella strains from a Saudi hospital. METHODS: From October 2015 to December 2016, a total of 149 Salmonella strains were collected from stool specimens from patients admitted to King Fahad Hospital of the University, AlKhobar, Saudi Arabia using CHROMagar Salmonella. The organism identification and antimicrobial susceptibility testing were performed using Vitek 2 system. Strain serogrouping was performed using Wellcolex color Salmonella kit. Fluoroquinolone resistance genes, extended-spectrum ß-lactamases (ESBLs), and AmpC ß-lactamase were determined using polymerase chain reaction (PCR). Enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) was used to determine clonal relatedness. RESULTS: The resistance rates to cefotaxime were 1.3% and ciprofloxacin 19.5%. Plasmid mediated quinolone resistance (PMQR) genes, qnrB and qnrS, were detected in 8 strains, qnrB (n=5) and qnrS (n=3), respectively. No ESBLs, AmpC, or mutations in the topoisomerases were detected. Salmonella isolates formed 7 clusters with similarity. CONCLUSIONS: This study reveals the emergence of fluoroquinolone resistant Salmonella in the region imposing public health concerns.

Molecular epidemiology of carbapenem-resistant Acinetobacter baumannii isolates in the Gulf Cooperation Council States: dominance of OXA-23-type producers.

The molecular epidemiology and mechanisms of resistance of carbapenem-resistant Acinetobacter baumannii (CRAB) were determined in hospitals in the states of the Cooperation Council for the Arab States of the Gulf (Gulf Cooperation Council [GCC]), namely, Saudi Arabia, United Arab Emirates, Oman, Qatar, Bahrain, and Kuwait. Isolates were subjected to PCR-based detection of antibiotic resistance genes and repetitive sequence-based PCR (rep-PCR) assessments of clonality. Selected isolates were subjected to multilocus sequence typing (MLST). We investigated 117 isolates resistant to carbapenem antibiotics (either imipenem or meropenem). All isolates were positive for OXA-51. The most common carbapenemases were the OXA-23-type, found in 107 isolates, followed by OXA-40-type (OXA-24-type), found in 5 isolates; 3 isolates carried the ISAba1 element upstream of blaOXA-51-type. No OXA-58-type, NDM-type, VIM-type, or IMP-type producers were detected. Multiple clones were detected with 16 clusters of clonally related CRAB. Some clusters involved hospitals in different states. MLST analysis of 15 representative isolates from different clusters identified seven different sequence types (ST195, ST208, ST229, ST436, ST450, ST452, and ST499), as well as three novel STs. The vast majority (84%) of the isolates in this study were associated with health care exposure. Awareness of multidrug-resistant organisms in GCC states has important implications for optimizing infection control practices; establishing antimicrobial stewardship programs within hospital, community, and agricultural settings; and emphasizing the need for establishing regional active surveillance systems. This will help to control the spread of CRAB in the Middle East and in hospitals accommodating transferred patients from this region.

Molecular characterization of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in the countries of the Gulf cooperation council: dominance of OXA-48 and NDM producers.

The molecular epidemiology and mechanisms of resistance of carbapenem-resistant Enterobacteriaceae (CRE) were determined in hospitals in the countries of the Gulf Cooperation Council (GCC), namely, Saudi Arabia, United Arab Emirates, Oman, Qatar, Bahrain, and Kuwait. Isolates were subjected to PCR-based detection of antibiotic-resistant genes and repetitive sequence-based PCR (rep-PCR) assessments of clonality. Sixty-two isolates which screened positive for potential carbapenemase production were assessed, and 45 were found to produce carbapenemase. The most common carbapenemases were of the OXA-48 (35 isolates) and NDM (16 isolates) types; 6 isolates were found to coproduce the OXA-48 and NDM types. No KPC-type, VIM-type, or IMP-type producers were detected. Multiple clones were detected with seven clusters of clonally related Klebsiella pneumoniae. Awareness of CRE in GCC countries has important implications for controlling the spread of CRE in the Middle East and in hospitals accommodating patients transferred from the region.