Detection of bla SHV and bla CTX-M genes in ESBL producing Klebsiella pneumoniae isolated from Egyptian patients with suspected nosocomial infections

The correct identification of the genes involved in ESBL mediated resistance is necessary for the surveillance and epidemiological studies of their transmission in hospitals. The aim of the present study was to find the prevalence of ESBL producing Klebsiella pneumonia among K. pneumonia isolates separated from Egyptian patients with suspected nosocomial infections, to detect their drug resistance pattern and to look for bla SHV and bla CTX-M genes in such organisms. 138 K. pneumonia isolates from Egyptian patients with suspected nosocomial infections were screened for ESBL production by the pattern of antimicrobial susceptibilities. Phenotypic identification for ESBL production was confirmed by double disc synergy test, phenotypic confirmatory double disc test and by MicroScan panel system. bla SHV and bla CTX-M genes in ESBL producing K. pneumonia were detected using multiplex PCR. The prevalence of ESBL producing K. pneumonia was 21% [30/138]. Their pattern of antimicrobial susceptibility showed that 90% was resistant to [Sulphamethoxazole / Trimethoprim], 70% was resistant to [Amoxicillin / Clavulanate], 63.3% was resistant to Cefotaxime and Ceftazidime, 60% was resistant to Amikacin, 46.7% was resistant to Doxycycline, Cefoxitin, Ceftriaxone and Levofloxacin, 40% was resistant to Cefepime, 20% was resistant to Ertapenem and [Sulbactam / Cefoperazone], 13.3% was resistant to [Piperacillin / Tazobactam], 10% was resistant to [Imipenem/Cilastatin] and Gentamycin and 6.7% was resistant to Meropenem and Ciprofloxacin. Among the ESBL producing K. pneumoniae, three out of 30 [10%] and 16 out of 30 [53.3%] were positive for bla SHV and bla CTX-M genes respectively. It could be concluded that ESBL producing isolates of K. pneumonia have been increasingly recognized in the hospital settings in Egypt and are associated with multiple drug resistance. Thus, molecular identification of the genes encoding beta lactamases would be essential for a reliable epidemiological investigation of their transmission in hospitals and antimicrobial resistance

DNA microarray for identification of fungal pathogens causing invasive fungal infections [IFIs] in neutropenic patients

Opportunistic invasive fungal infections [IFIs] remain as important cause of morbidity and mortality. Candida and Aspergillus species are the most common fungi that cause disease in immunocompromised patients and transplant recipients. This study was designed to identify Candida and AspergilIus spp. as possible causes of IFIs in neutropenic patients with different hematological diseases, using high multiplexing capacity of DNA microarray [species identification array]. Twenty eight patients admitted to Hematology unit-Ain Shams University Hospitals with provisional diagnosis of lFl were enrolled in this study. Venous blood samples were collected to detect Candida and Aspergillus spp. using DNA microarray. Nineteen out of 28 studied patients [67.9%] were infected with Candida and Aspergillus spp. Invasive aspergillosis constituted 13/19[68.4%] distributed as follows: A.fumigatus 6/19 [31.6%], A.flavus 4/19 [21%] and A.niger 3/19 [15.8%]. On the other hand, Fl with Candida spp. constituted 6/19 [31.6%] distributed as follows; C.glabrata 3/19 [15.8%], C.tropicalis 2/19 [10.5%] and C.albicans 1/19 [5.3%]. Duration of hospital stay [mean +/- SD = 30.8 +/- 4 days] was statistically significant among the infected group in comparison to other patients [mean +/- SD = 22.7 +/- 2.3 days]. Nineteen out of 28 studied patients [67.9%] were infected with Candida and Aspergillus spp. Invasive aspergillosis constituted 13/19 [68.4%], while Candidal infection constituted 6/19 [31.6%]. DNA microarray represents a reliable method of potential use in clinical laboratories for parallel one-shot detection and identification of the most common fungal pathogens at the species level for prompt management of infection with tailored antifungal treatments