Prevalence of extended-spectrum ß-lactamase (ESBL) and molecular detection of blaTEM, blaSHV and blaCTX-M genotypes among Enterobacteriaceae isolates from patients in Khartoum, Sudan.

INTRODUCTION: the emergence of antibiotic resistance pathogens is an important health risk. Usually Gram negative bacteria acquire resistance to beta-lactam antibiotics by beta-lactamase production. The objectives of this study was to assess the prevalence of ESBL and to detect the frequency of blaTEM, blaSHV and blaCTX-M genotypes among ESBL producing Enterobacteriaceae isolates from patients in Khartoum, Sudan. METHODS: a total of 171 isolates of Enterobacteriaceae were recovered from hospitals in Khartoum, Sudan (2014 -2015) were used to detect ESBL production using disc diffusion method. blaTEM, blaSHV and blaCTX-M genes were investigated by PCR based methods using gene-specific primers. RESULTS: the high resistance among Enterobacteriaceae was noticed in ciprofloxacin (72%) and ofloxacin (73%). ESBL production was mainly in Escherichia Coli (38%) and Klebsiella pneumonia (34%). Prevalent genotypes were blaTEM (86%), blaCTX-M (78%) and blaSHV (28%). These were found mainly in Escherichia Coli (38%, 37%, 2%) and K. pneumonia (34%, 31%, 26.1%). The majority of ESBL producing isolates possess more than one ESBL genes. CONCLUSION: the ESBL production in Enterobacteriaceae was high, with blaTEM and blaCTX-M genotypes more prevalent. Public health and laboratory standard of excellence is needed to reducing the spread of resistant pathogens.

Coagulase gene polymorphisms of Staphylococcus aureus isolates from patients at Kosti Teaching Hospital, Sudan.

BACKGROUND: Staphylococcus aureus is a common cause of nosocomial infections. Epidemiological typing of S. aureus enables control of its spread. The objective of this study was to investigate coagulase gene polymorphisms of S. aureus isolated from patients at Kosti Hospital in Sudan. METHODS: In total, 110 S. aureus isolates were recovered from 110 patients who were enrolled in the study. S. aureus strains were isolated on blood agar and MacConkey agar and then identified by conventional tests. Resistance to methicillin was determined by detection of the mecA gene. Polymorphism in the coagulase gene (coa) was investigated using PCR followed by AluI RFLP analysis. RESULTS: Methicillin-resistant S. aureus accounted for 62/110 (56 %) of the isolates. PCR of the coa gene showed two different amplicons, one of 680 bp detected in 83/110 (75.5 %) of the isolates and one of 790 bp detected in 27/110 (24.5 %). When digested with the AluI enzyme, the 790 bp amplicon resulted in three restriction fragments of 500, 210 and 80 bp (coa1). Restriction of the 680 bp amplicon gave two patterns; the first (coa2) was found in 22/110 (20 %) of the isolates and showed four fragments of 210, 210, 180 and 80 bp, and the second (coa3) was found in 61/110 (55.5 %) and revealed three fragments of 390, 210 and 80 bp. Most of the coa3 isolates (75.4%) were methicillin-resistant. CONCLUSION: Three polymorphic genotypes of S. aureus were identified in patients at Kosti Hospital. The coa3 genotype was the predominant one and was mostly detected in methicillin-resistant isolates.

The impact of cathelicidin, the human antimicrobial peptide LL-37 in urinary tract infections.

BACKGROUND: The defense mechanisms of the urinary tract are attributed mainly to the innate immune system and the urinary tract urothelium which represent the first line of defense against invading pathogens and maintaining sterility of the urinary tract. There are only a few publications regarding cathelicidin (LL-37) and a urinary tract infection (UTI). This study was done to investigate the plasma and urine levels of human LL-37 in patients with UTI. METHODS: A case-control study was conducted at Omdurman Hospital, Sudan during the period from August 2014 to May 2017. The cases were patients with confirmed UTI and the controls were healthy volunteers without UTI. Sociodemographic and clinical data were obtained from each participant using questionnaires. Urine cultures and antimicrobial susceptibility were tested. Plasma and urine levels of LL-37 were determined using an enzyme-linked immunosorbent assay (ELISA) kit. SPSS (version 16.0) was used for analyses. RESULTS: Cases and controls (87 in each arm) were matched according to their basic characteristics. Compared with controls, the median (inter-quartile) LL-37 level in plasma [2.100 (1.700-2.700) vs. 1.800 (1.000-2.200) ng/ml, P = 0.002] and in urine [0.900 (0.300-1.600) vs. 0.000 (0.000-1.000) ng/mg creatinine, P < 0.001] was significantly higher in cases. There was no significant difference in the median plasma [2.1 (1.7-2.9) vs. 2.000 (1.700-2.400) ng/ml, P = 0.561] and urine [0.850 (0.275-2.025) vs. 0.900 (0.250-1.350) ng/mg creatinine, P = 0.124]. The uropathogenic Escherichia coli (UPEC) was the predominant isolate, n = 38 (43.7%). LL-37 levels between the E. coli isolates and the other isolated organisms. There was no significant correlation between plasma and urine LL-37 levels (r = 0.221), even when the data of the cases were analyzed separately. CONCLUSION: LL-37 is notably increased among patients with UTI compared with normal control subjects. Severity of UTI increases the levels of LL-37. The increased level was not only in the patient's urine, but has also been observed in the patient's plasma. Detection of increased levels of LL-37 could help to differentiate subjects with suspected UTI. Accordingly, LL-37 could act as a good marker for diagnosing UTIs.

Assessment of methicillin resistant Staphylococcus Aureus detection methods: analytical comparative study.

INTRODUCTION: The heterogeneous expression of methicillin resistance in Staphylococcus aureus (MRSA) affects the efficiency of tests available to detect it. The objective of this study was to assess four phenotypic tests used to detect MRSA. METHODS: This is an analytical comparative study conducted among sudanese patients during period from May 2012 to July 2014, Staphylococcus aureus strains were isolated and identified by conventional methods, and then confirmed by PCR detection of coagulase gene. PCR detection of mecA gene was used as a gold standard to assess oxacillin resistance screen agar base (ORSAB), oxacillin disc, cefoxitin disc (at different temperatures and incubation periods) and MRSA-latex agglutination test. S.aureus ATCC 25923 was used as control. Sensitivity and specificity were calculated. RESULTS: MRSA- latex agglutination was the most accurate test; it showed 100% of both sensitivity and specificity, followed by cefoxitin disc with sensitivity of 98.48% and specificity of 100%. However, both of oxacillin disc and oxacillin resistance screen agar base showed less accurate results, and were affected by incubation periods. Oxacillin disc after 24 h incubation both at 30°C and 35°C showed sensitivity and specificity values of 87.88% and 96.23%, respectively. However, after 48h incubation the test at 30°C showed sensitivity and specificity values of 89.39%, and 94.34%, respectively. At 35°C (48h) it showed values of 89.39%, 92.45% respectively. Specificity of ORSAB was more than oxacillin disc at 35°C after 24h incubation 98.11% and 96.23%, respectively. CONCLUSION: MRSA- latex agglutination and cefoxitin disc diffusion tests are recommended for routine detection of MRSA.

Detection and sequencing of rotavirus among sudanese children.

INTRODUCTION: Diarrheal diseases are a big public health problem worldwide, particularly among developing countries. The current study was conducted to detect and characterize group A rotavirus among admitted children with gastroenteritis to the pediatric hospitals, Sudan. METHODS: A total of 755 stool samples were collected from Sudanese children with less than 5 years of age presenting with acute gastroenteritis during the period from April to September 2010. Enzyme-linked immunosorbent assay (ELISA) was used to Detection of Rotavirus antigens. Ribonucleic acid (RNAs) were extracted from rotavirus-positive stool samples using (QIAamp® Viral RNA Mini Kit). (Omniscript® Reverse Transcription kit) was used to convert RNA to complementary Deoxyribonucleic acid (cDNA). The cDNAs were used as template for detection of VP4-P (P for Protease-sensitive) and VP7-G (G for Glycoprotein) genotyping of Rotavirus using nested PCR and sequencing. RESULTS: Out of the 755 stool samples from children with acute gastroenteritis, 121 were positive for rotavirus A. Among 24 samples that were sequenced; the VP7 predominant G type was G1 (83.3%), followed by G9 (16.7%). Out of these samples, only one VP4 P[8] genotype was detected. CONCLUSION: As a conclusion the VP7 predominant G type was G1, followed by G9 whereas only one VP4 genotype was detected and showed similarity to P[8] GenBank strain. It appears that the recently approved rotavirus vaccines in Sudan are well matched to the rotavirus genotypes identified in this study, though more studies are needed.

Antimicrobial susceptibility and molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacteriaceae from intensive care units at Hamad Medical Corporation, Qatar.

BACKGROUND: The emergence of extended-spectrum beta-lactamase (ESBL)-producing isolates has important clinical and therapeutic implications. High prevalence of ESBL-producing Enterobacteriaceae has been reported in the literature for clinical samples from a variety of infection sites. The present study was undertaken to evaluate the prevalence of ESBL-producing Enterobacteriaceae, and to perform molecular characterization and antimicrobial susceptibility testing of clinical isolates from patients admitted to the intensive care units at Hamad Medical Corporation, Doha, Qatar, from November 2012 to October 2013. METHODS: A total of 629 Enterobacteriaceae isolates were included in the study. Identification and susceptibility testing was performed using Phoenix (Becton Dickinson) and the ESBL producers were confirmed by double-disk potentiation as recommended by the Clinical and Laboratory Standards Institute. Molecular analysis of the ESBL producers was performed by polymerase chain reaction. RESULTS: In total, 109 isolates (17.3 %) were confirmed as ESBL producers and all were sensitive to meropenem in routine susceptibility assays. Most of the ESBL producers (99.1 %) were resistant to amoxicillin/clavulanic acid and ceftriaxone and 93.6 % were resistant to cefepime. Among the ESBL-producing genes, bla CTX-M (66.1 %) was the most prevalent, followed by bla SHV (53.2 %) and bla TEM (40.4 %). CONCLUSIONS: These findings show the high prevalence of ESBL-producing Enterobacteriaceae within the intensive care units at Hamad Medical Corporation, Qatar, and emphasize the need for judicious use of antibiotics and the implementation of strict infection control measures.

Rotavirus infection among Sudanese children younger than 5 years of age: a cross sectional hospital-based study.

INTRODUCTION: In Sudan, rotavirus has been one of the important causative agents of diarrhea among children. Rotavirus A is well known as the leading cause of diarrhea in young children worldwide. It was estimated to account for 41% of hospitalized cases of acute gastroenteritis among children in Sub-Saharan Africa. This study aimed to determine the prevalence and the common clinical presentations of rotavirus A infection among Sudanese children with gastroenteritis seeking management in hospitals. METHODS: 755 Sudanese children less than 5 years of age suffering from acute gastroenteritis in hospital settings were included. The positive stool specimens for rotavirus A was used for extract Ribonucleic acid (RNA) and the RNA product was loaded on formaldehyde agarose gel and visualized under UV illumination. RESULTS: Of the 755 children, 430(57%) were males while 325(43%) were female. The age of children ranged from 1 to 60 months. There were 631 (84%) children who were less than 24 months of age. Out of the 755 stool samples, 121(16%) were positive for rotavirus. Of the 121 infected children with rotavirus, 79(65.3%) were male and 42(34.7%) were female and the highest infection rate was seen among 91(75.2%) of children up to 12 months of age. Children of illiterate parents were more infected with rotavirus than children of educated parents. Severe dehydration present among 70% of infected children with rotavirus. CONCLUSION: Since this study is hospital-based, the 16% prevalence rate may not reflect the true prevalence among Sudanese children, thus a community-based surveillance is needed.

International spread of major clones of methicillin resistant Staphylococcus aureus: nosocomial endemicity of multi locus sequence type 239 in Saudi Arabia and Romania.

Phenotypically identified methicillin resistant Staphylococcus aureus (MRSA) strains from several hospitals in Romania and Saudi Arabia (n = 103 and 68, respectively) were confirmed to be MRSA by mecA PCR and PBP-2' based latex agglutination. Subsequently, strains were differentiated at the sub-species level using pulsed field gel electrophoresis (PFGE) of SmaI DNA macro-restriction fragments. Comparison of the PFGE fingerprints identified major clusters of strains, persistently present in the various hospitals. Endemicity of certain strains was identified, amongst others one due to a particularly methicillin resistant type in the burn wound sector of the Romanian hospital. No PFGE-based overlap was found between the Saudi and Romanian strains. However, multi locus sequence typing (MLST), performed for 20% of all strains, revealed that genuine genetic similarity was obscured by the PFGE analysis. In both the Romanian and Saudi hospitals the renowned sequence type (ST) 239 was very over-represented. This was especially apparent in Saudi Arabia, where all strains except two shared the ST 239 genotype. This clonal type has previously been identified in a variety of other countries. Despite the MLST concordance, PFGE data indicate that ST 239 diversifies while maintaining its core genome intact. ST 80, another previously but less frequently identified clone, was introduced in 2000 in the Romanian institutes and persisted over the past 3 years as a frequent cause of infections in a surgical department. The successful MRSA types can acquire prominent positions in hospitals of previously low-endemicity MRSA status.

Endemic nosocomial infections and misuse of antibiotics in a maternity hospital in Saudi Arabia.

Patients admitted during a 6-month period to a maternity hospital in Saudi Arabia were studied for nosocomial infections and misuse of antibiotics. Patient history and diagnosis on admission and subsequent clinical and laboratory data were analysed. Infection developing from 72 h after admission was considered nosocomial. Therapeutic and prophylactic data as recorded on the patients' charts were assessed for possible misuse of antibiotics. Of 3439 patients, 136 (4.0%) developed nosocomial infection: 2.0%, 8.9% and 37.7% in obstetric, gynaecologic and nursery patients, respectively. Infections among adults were mostly found in the urinary (44.4%) and lower genital (33.3%) tracts. Among newborns, over 70% of cases were eye and ear (29.8%), skin (26.2%) and blood (19.0%) infections. Gram-negative bacteria caused 65.7% of the infections. Over 90% of the bacterial isolates were multidrug-resistant. About 24% of patients received single or multiple antibiotics; 57.2% were misused. The minimal hospital cost estimate for both nosocomial infections and misused antibiotics was US $318,705. The findings of this study, the first of its type in this region, should prompt improved infection control measures as well as educational and antibiotic restriction interventions.