Characterization of Carbapenemase-Producing Enterobacteriaceae from Patients in Amman, Jordan.

We sought to detect and determine the genetic diversity of carbapenemase-producing Enterobacteriaceae (CPE) isolated from clinical specimens in Amman, Jordan. From five hospitals, a total of 2,759 isolates had antimicrobial susceptibilities determined via Vitek II, of which 28 (1%) were carbapenem resistant. Species identifications were determined via matrix-assisted laser desorption ionization time-of-flight mass spectrometry and carbapenemase gene detection via real-time PCR indicated that 23 (82.1%) isolates were Klebsiella pneumoniae (OXA-48-like, n = 7; NDM, n = 14; OXA-48-like and NDM, n = 2), four (14.2%) were Enterobacter cloacae complex (NDM, n = 3 and VIM, n = 1), and one (3.5%) was Escherichia coli (NDM). Sequencing of carbapenemase gene amplicons from a subset of isolates identified blaNDM-1, blaOXA-48, and blaVIM-4 alleles. Strain typing detected seven different K. pneumoniae variable number tandem repeat types, consistent with mostly sporadic occurrences along with limited clonal spread. E. cloacae complex isolates were diverse by pulsed-field gel electrophoresis, with a maximum relatedness of 70%. Plasmid restriction fragment length polymorphism (pRFLP) revealed four distinct profiles associated with NDM-encoding plasmids that were positive for replicons of the FII(K)/FIB or FIB incompatibility (Inc) groups via PCR-based replicon typing. OXA-48-encoding IncL/M plasmids differed by two pRFLP bands. The results show diverse CPE produce OXA-48 and NDM-1 enzymes in Jordan and that the carbapenemase genes are distributed on diverse plasmids in Jordanian hospitals, with some limited evidence for related clusters occurring, emphasizing the need for strict infection control measures.

Molecular characterization of the circulating Bacillus anthracis in Jordan.

To understand the biomolecular charcteristics of Bacillus anthracis in Jordan, 20 blood smear slides from dead animals with suspected anthrax were analyzed using conventional and molecular approaches. All slides were positive for B. anthracis by conventional staining but no growth of the organism on selective media was detected. However, of the 20 samples, 16 were B. anthracis DNA-positive using polymerase chain reaction (PCR). Seven samples provided enough quantity and quality of DNA, and their multilocus variable tandem repeat analysis (MLVA)-15 loci analysis revealed two different genotypes. All genotypes were belonging to A.B..r. 008/009 which is very common in Asia and Europe. Single nucleotide repeat (SNR) analysis revealed that there were no sub genotypes. Molecular diagnosis of animal anthrax in Jordan is not used routinely; henceforth, official diagnosis of anthrax is based on the observation of the slides by optical microscope and this can often cause reading errors. Therefore, the prevalence of the disease in Jordan might be slightly lower than that reported by the official bodies.

Rare occurrence of mupirocin resistance among clinical Staphylococcus isolates in Jordan.

Staphylococcal infections have high occurrence in Jordanian patients. This study was carried out to determine the rates of high- and low-level mupirocin resistance (MupH and MupL) among staphylococci with the molecular characterization. Two hundred and thirty-two non-duplicate Staphylococcus spp. isolated from different clinical specimens were tested for mupirocin susceptibility using disk diffusion method and minimum inhibitory concentration (MIC). Resistance genes and clone relatedness was studied using polymerase chain reaction (PCR) and enterobacterial repetitive intergenic consensus primers (Eric-PCR) for the latter. Plasmid curing was performed to determine the genetic location of MupA gene. Among the 232 strains, 144 (62%) were methicillin-resistant Staphylococcus aureus (MRSA), 33 (14.2%) methicillin-susceptible Staphylococcus aureus (MSSA) and 55 (23.7%) were of other coagulase-negative Staphylococcus spp. (CoNS). Of all strains tested, only 6 (2.6%) were mupirocin resistant. MecA gene was detected in both MupL and MupH strains but MupA gene was only detected in MupH. Plasmid curing improved the plasmidic location of MupA gene. Molecular typing by Eric-PCR method revealed heterogenicity of the genetic make up of our MupL and MupH strains. Staphylococci with MupA-carrying genes are present in Jordanian hospitals, but thank to the limited use of mupirocin, they remain rare.