Dalia Kadry Ismail

Detection of viral acute lower respiratory tract infection in hospitalized infants using real-time PCR

Abstract Introduction Acute lower respiratory tract infection in children causes significant morbidity in the developing countries. Documentation of virus infection using PCR and clinical characteristics of patients affected with viral pneumonia are reviewed in this study. Methods 51 children less than three years admitted to the Pediatric Hospital, Cairo University with viral pneumonia were included. All patients had undergone nasopharyngeal aspirate for PCR viral detection. Results A total of 51 cases were enrolled in the study, of which 7 cases were negative while 44 children were positive for viruses. The most common respiratory virus was Rhinovirus in 32 patients (72.2%), then parainfluenza virus (PIV) in 12 (27.3%), of which subtypes PIV1 were 2 (4.5%), PIV3 were 5 (11.4%) and PIV4 were 5 (11.4%) cases. The third common viruses were respiratory syncytial virus (RSV) in 9 (20.5%) cases of which 3 (6.8%) were RSVA and 6 (13.6%) were RSVB and adenovirus in 9 cases (20.5%). Boca virus was found in 8 (18.2%) patients, corona virus 2 (4.5%) patients, H1N1 2 (4.5%) patients, enterovirus 2 patients (4.5%) and human metapneumovirus in one case (2.3%). Influenza B and PIV2 were not detected. Coinfection was found in 28 (63.7%). Mortality occurred in 12 (23.5%). There was no significant relation between virus type or coinfection with disease severity. Conclusions RV was the most commonly detected virus in children under 3years admitted with acute lower respiratory tract infections. Coinfection was present in the majority of our patients; however it was not related significantly to parameters of disease severity.

Diabetic Foot Infection : Microbiological Causes with Special Reference to Their Antibiotic Resistance Pattern

Background: Foot infections are one of the major complications of diabetes mellitus and are a significant risk factor for lower extremity amputation. Providing effective antimicrobial therapy is an important component in treating these infections. Objectives: This study investigates the microbial causes of diabetic foot infections and their antibiotic susceptibility pattern. Methodology: A prospective study of 80 patients with diabetic foot infections admitted to Cairo university hospitals was undertaken. Samples were collected by using sterile swabs and they were processed. Detailed antimicrobial susceptibility testing was performed according to CLSI guidelines 2014, including testing for Methicillin Resistant Staphylococcus aureus (MRSA), ESBL and carbapenemase production. Results: In the present study, a single organism was isolated from 48.75% patients and mixed bacterial growths were seen in 51.15% patients. Gram- negative bacteria (56.08%), were more commonly isolated compared with Gram-positive bacteria (27.7%), while gram-negative anaerobes accounted for (8.1%),and Candida species (8.1%). The commonest isolate was Proteus mirabilis (16.8%) followed by Escherichia coli (13.5%), Methicillin Sensitive Staphyloccus aureus (MSSA) (11.4%), Pseudomonas spp (10.8%), and MRSA (10.1%). Among the gram-negative bacteria 49% were ESBL producers and 3.6%were carbapenemases producerswhile46.8% Staphylococcus aureus were methicillin resistant Staphylococcus aureus. Overall 42.5% isolates were identified as MDR. Vancomycin was found to be the most effective against Gram-positive bacteria, whereas imipenem, amikacin and colistin were most effective against Gram-negative bacteria. Conclusion: This study showed a preponderance of gram negative bacilli among the isolates from the diabetic foot ulcers. Knowledge on the antibiotic sensitivity pattern of the isolates will be helpful in determining the drugs for the empirical treatment of diabetic ulcers

Integron-mediated multidrug resistance in extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolated from fecal specimens in Egypt.

BACKGROUND The increasing incidence of hospital-acquired infections due to extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae represent a major health problem because of few therapeutic alternatives. The fecal flora can represent a reservoir for ESBL genes. Integrons are genetic structures capable of capturing gene cassettes that usually encode antibiotic-resistance determinants. OBJECTIVES To investigate the antimicrobial susceptibility of fecal isolates of ESBL-producing Escherichia coli and Klebsiella pneumoniae from hospitalized and nonhospitalized Egyptian patients and to determine the prevalence of class 1 and class 2 integrons together with the most common ESBL-producing genes (bla TEM, SHV, CTX-M, and OXA) among the collected isolates. MATERIALS AND METHODS Ninety-six fecal samples were collected: 48 samples from hospitalized patients admitted at Kasr Al-Ainy University Hospital, Cairo and 48 from outpatient clinics. Samples were inoculated on MacConkey agar and identified. All isolates were tested for their susceptibility to different antimicrobial agents using a standard disk diffusion method. The double-disk synergy test was applied for screening ESBL. All ESBL-producing isolates were confirmed by molecular testing to detect ESBL-encoding genes (SHV, TEM, CTX-M, and OXA). To identify the strains carrying integrons 1 and 2, the conserved regions of integron-encoded integrase gene intI1 and intI2 were amplified. RESULTS E. coli isolates accounted for 52.1% of the isolates collected from hospitalized patients and 60.4% of those collected from outpatient clinics. Results of the double-disk synergy test were positive in all E. coli and K. pneumoniae isolates, indicating the presence of ESBL production. Isolates of both groups showed variably high degrees of resistance to ciprofloxacin and co-trimoxazole. The most predominant ESBL gene in both groups was the bla CTX-M gene (93.8%) and the least prevalent was the bla OXA gene, which was not detected in any of the study isolates. Between the other two genes, the bla TEM gene was more common than the bla SHV gene in the two study groups. Class 1 integron was more prevalent among hospitalized patients, being detected in 64.6% of isolates from this group. Class 1 integron was linked with the bla CTX-M gene (P=0.039). Class 2 integron was more prevalent in the nonhospitalized group (85.4%) compared with the hospitalized group (50%) (P<0.001). CONCLUSION AND RECOMMENDATIONS ESBL-producing E. coli and K. pneumoniae showed a marked degree of antibiotic resistance in both hospitalized and nonhospitalized study groups. The high prevalence of class 1 and 2 integrons among isolates of both groups has a serious impact on the spread of antibiotic resistance.

Pneumococcal infection among hospitalized Egyptian children.

AIM We aimed to describe the detection rate spectrum of clinical manifestations, and outcome of pneumococcal disease in children younger than 5 years admitted to the largest referral pediatric hospital in Egypt. MATERIALS AND METHODS This was a hospital-based study to detect laboratory-confirmed Streptococcus pneumoniae cases among children younger than 5 years. Data on demographic characteristics, clinical diagnosis, comorbidities, diagnostic tests, antibiotic resistance, and clinical outcome were collected during the study years from 2008 to 2011. RESULTS During the 4-year study period, 22 018 cases younger than 5 years had cultures performed at Cairo University Pediatric Hospital microbiology laboratory. We estimated the annual detection rate of total Streptococcus pneumonia infection to be 54.5/100 000. The incidence of invasive pneumococcal disease (IPD) was half the incidence of non-IPD (18.2 and 36.4/100 000, respectively). Infants of 1 year or younger were statistically more vulnerable to Streptococcus pneumonia infection compared with children between 1 and 5 years of age (annual rate: 110.5/100 000 and 21.6/100 000, respectively). The overall pneumococcal annual case fatality was 33.3% and was higher in IPD (75%) than in non-IPD (12.5%) cases. There was an obviously increasing trend of the pneumococcal detection rate throughout the 4 years of the study (P<0.0001). CONCLUSION AND RECOMMENDATIONS Our results confirm the substantial and increasing pneumococcal infection, the emerging of multidrug resistant isolates, and the vulnerability of the younger age group and high-risk population, which calls for a national surveillance to inform policy and decision-making before national wide vaccine introduction.

Reduced susceptibility of Enterococcus spp. isolates from Cairo University Hospital to tigecycline: Highlight on the influence of proton pump inhibitors

OBJECTIVES The incidence of reduced susceptibility to tigecycline (TIG) is increasing. This study aimed to analyse the in vitro activity of TIG against Enterococcus spp. isolates recovered from hospitalised patients and to evaluate the effect of omeprazole on the in vitro antimicrobial activity of TIG against several enterococcal species. METHODS A total of 67 Enterococcus clinical isolates were identified by MALDI-TOF/MS and multiplex PCR. Minimum inhibitory concentrations (MICs) of TIG alone and in combination with omeprazole (10, 30 and 60mg/L) were determined by broth microdilution. Antibiotic susceptibility to other antibiotics was determined by disk diffusion. The presence of van, tet(X) and tet(X1) genes was tested by multiplex PCR. RESULTS Of the 67 Enterococcus isolates, 2 (3.0%) were resistant to TIG and 13 (19.4%) were intermediate-resistant according to EUCAST. The frequencies of resistance to norfloxacin (80.6%), doxycycline (80.6%), levofloxacin (74.6%) and ciprofloxacin (71.6%) were highest, whilst that of vancomycin (25.4%) was lowest. The vanA gene was detected in 11 Enterococcus isolates (8 Enterococcus faecalis, 3 Enterococcus faecium), vanB in 3 Enterococcus isolates (2 E. faecium, 1 E. faecalis) and vanC-2/3 in 3 Enterococcus casseliflavus. Nine isolates (13.4%) were positive for tet(X1). TIG resistance occurred both in patients receiving or not TIG and/or omeprazole. Omeprazole increased TIG MICs by 4-128-fold. CONCLUSIONS The possibility of selection of TIG-non-susceptible Enterococcus in the gut may occur with long-term use of omeprazole. Omeprazole influenced TIG activity in a concentration-dependent manner. To our knowledge; this is the first report of TIG-non-susceptible Enterococcus spp. in Egypt.

A study of biofilm on endotracheal tubes in pediatric intensive care unit

Ventilator-associated pneumonia (VAP) is one of the most common hospital-acquired infections occurring in mechanically ventilated patients and is associated with increased mortality, longer pediatric intensive care unit (PICU) stay, and health-related costs [1]. Due to the role of endotracheal tubes (ETTs) in the pathogenesis of VAP, some authors suggest that it should be renamed ETT-associated pneumonia [2]. This study was performed to evaluate the existence of biofilm in the ETT) of mechanically ventilated children, and to study a microbial link between biofilm flora and bacteria causing VAP. Biofilms have public health significance owing to their role in certain infectious diseases and their role in a variety of device-related infections [3]. This was a prospective study conducted on 20 children with ETT who were evaluated for biofilm existence using scanning electron microscopy. A total of 20 children were enrolled in the study. Of them, 17 (85%) children showed biofilm formation on the luminal surface of ETT (Figs. 1

Emergence of Gram-Negative Bacilli with Concomitant bla NDM-1 - and bla OXA-48 -Like Genes in Egypt

Multidrug-resistant Gram-negative organisms have emerged as a major threat to hospitalized patients, and are associated with serious morbidity and mortality. This study aimed to characterize carbapenem resistance genes among Gram-negative bacilli isolated from clinical samples from patients in the intensive care unit of Cairo University Hospital. A total of 211 samples were collected from patients showing clinical evidence of infection. Bacteria were isolated and identified by conventional microbiological methods. Acinetobacter baumannii isolates were furtherly characterized by polymerase chain reaction (PCR), using primers specific for blaOXA-51-like genes. The Kirby Bauer disc diffusion method was used to determine susceptibility patterns of isolates, and carbapenem resistance was further examined by a modified Hodge test. Positive isolates were tested for the presence of blaKPC, blaOXA-48, and blaNDM-like genes by PCR. NDM gene types were determined by direct sequencing. From the 211 samples, 229 Gram-negative bacilli were isolated. Fifty isolates (21.2%) were resistant to carbapenem. PCR analysis showed that none of the 50 isolates carried blaKPC-like genes, while 24 (48%) isolates carried blaOXA-48-like genes, 8 (16%) carried blaNDM-1, and five isolates (10%) carried both blaNDM-1 and blaOXA-48-like genes. These results indicate that continuous surveillance of these multidrug-resistant pathogens is urgently required. And that is very important is to activate the antimicrobial stewardship programs of which the most important is restriction of the big gun antibiotics like carbapenems, colistin, tigecyclin and vancomycin and restricting their prescription to privileged specialties.

Evaluation of a novel commercial quaternary ammonium compound for eradication of Mycobacteria, HCV and HBV in Egypt

Endoscopes are a common source of outbreaks of healthcare-associated infections. It is therefore important to identify high-level disinfectants capable of eliminating or killing all vegetative bacteria, mycobacteria, and viruses. Aldehydebased disinfectants are most commonly used in clinical practice but resistance has recently been detected and side effects associated with these disinfectants are well documented. In this study, we evaluated Virusolve+® EDS, a novel quaternary ammonium compound formulation supplied by Amity international, against Mycobacterium bovis (ATCC-27289), hepatitis C virus (HCV)-positive serum and hepatitis B surface antigen-positive serum. We also compared its efficacy against Cidex® (glutaraldehyde 2%), an aldehyde-based disinfectant. M. bovis showed no growth after 10 weeks with either Virusolve+® or Cidex®. Virusolve+® achieved a 104- fold reduction in the initial 106 HCV load under clean conditions (without red blood cells) for 20 min, whereas Cidex® achieved this reduction under clean and dirty conditions (without and with red blood cells, respectively) after both 10 and 20 min. Both Virusolve+® and Cidex® were able to eradicate hepatitis B virus (HBV) infectivity under clean conditions after 10 and 20 min, whereas under dirty conditions they were only able to eradicate virus infectivity after 20 min. Virusolve+® EDS when compared with Cidex® showed equal mycobactericidal activity completely eradicating M. bovis. However, both showed comparable virucidal activity against HBV, which was more effective under clean conditions, emphasizing the importance of the cleaning step in endoscope reprocessing. Cidex® was more effective at eradicating HCV under dirty conditions after a short contact time.