ABSTRACT
OBJECTIVE: Carbapenemase-producing, or carbapenem-resistant, Enterobacteriaceae are an emerging threat to human and animal health because they are resistant to many of the last-line antimicrobials available for treatment of infection. The aim of this study was to analyse the antimicrobial resistance patterns and their encoding genes of Proteus mirabilis isolated in Constantine, Algeria. METHODS: A total of 108 Proteus, Morganella and Providencia (PMP) strains were isolated from a large variety of clinical specimens at University Hospital of Constantine in Algeria. Isolates were identified using the API 20E system and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Diagnostic accuracy was determined by independent comparison of each method to phylogenetic analysis based on 16S rRNA gene sequencing. Antimicrobial susceptibility was determined by the standard disk diffusion and Etest methods. The presence of antimicrobial resistance genes was screened for by PCR amplification and sequencing. RESULTS: A total of 72 PMP strains were multidrug-resistant (MDR). Among them, one P. mirabilis isolate was resistant to imipenem with a minimum inhibitory concentration (MIC) of ≥12µg/mL. PCR and sequencing showed the presence of various antimicrobial resistance genes, including blaCTX-M-15, blaTEM-1, blaTEM-2, blaPER-1, blaSHV-11, aadA1, aadA2, armA, aac(6')-Ib, aac(6')-Ib-cr, aac(3)-Ia and ant(2â³)-I, forming different resistance profiles. Moreover, the blaOXA-24 gene was detected in the imipenem-resistant P. mirabilis strain. CONCLUSION: In this study, a MDR P. mirabilis isolate harbouring the blaOXA-24, armA 16S rRNA methylase and aac(6)-Ib-cr genes was found for the first time in Algeria.
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Methyltransferases/genetics , Proteus mirabilis/drug effects , Proteus mirabilis/genetics , beta-Lactamases/genetics , Algeria , Bacterial Proteins/genetics , Humans , Microbial Sensitivity Tests , Proteus Infections/microbiology , Proteus mirabilis/enzymologyABSTRACT
Onychomycosis (OM) is a nail infection caused mainly by dermatophyte species but other species of yeast and moulds are frequently involved as well. Classical diagnosis has limitations thus empirical treatment is common. The usefulness of different real time PCR (RT-PCR) assays for identifying species causing OM was assessed in samples from seventy patients and fifteen controls. Conventional methods and four different RT-PCR assays were used: a panfungal, a pandermatophyte and two specific assays for detecting Candida spp. and Aspergillus spp. Fungal elements were visualised in 58% of the samples, and 54% of cultures were positive. Panfungal and pandermatophyte RT-PCR were positive in 28% and 60%, respectively, and the sensitivity relative to positive cultures was 47% and 90%. Candida spp. were detected in 76% of samples analysed and Aspergillus spp. in 60%. These species were also present in 80% of control cases. In conclusion, molecular techniques were useful but showed limitations. The panfungal assay showed a low sensitivity, the pandermatophyte assay was sensitive and specific but did not allow for differentiation among species of dermatophytes. Finally, the role of non-dermatophyte species detected by using specific RT-PCR techniques should be carefully analysed as these species were also present in healthy nails.
Subject(s)
Arthrodermataceae/isolation & purification , Fungi/isolation & purification , Onychomycosis/microbiology , Real-Time Polymerase Chain Reaction/methods , Trichophyton/isolation & purification , Arthrodermataceae/genetics , Candida/genetics , Candida/isolation & purification , DNA, Fungal/analysis , Female , Fungi/genetics , Humans , Male , Molecular Diagnostic Techniques , Nails/microbiology , Onychomycosis/diagnosis , Sensitivity and Specificity , Spain , Trichophyton/geneticsABSTRACT
Cutaneous leishmaniasis is endemic in Algeria, but clinical and parasitological data from this area are scarce. In order to document the transmission of this disease in a peri-urban setting, cutaneous lesions from patients living in Constantine City and surrounding areas were spotted on filter paper for diagnosis and species identification using real-time PCR. Surprisingly, Leishmania tropica was detected in 6/69 patients, and confirmation was obtained by sequencing. This observation suggests a modification of the epidemiology of cutaneous leishmaniasis in Algeria and should alert physicians and policy-makers to the risk of antimony treatment failure with this species.
Subject(s)
Leishmania tropica/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Algeria , Animals , DNA, Protozoan/analysis , Diagnosis, Differential , Humans , Leishmaniasis, Cutaneous/diagnosis , Polymerase Chain Reaction/methods , Rural Health , Urban HealthABSTRACT
We evaluated the use of real-time PCR for the identification of cutaneous Leishmania species. The assay, based on the melting curve analysis of fluorescent products, allows the discrimination of four culture adapted strains (L. infantum MHOM/TN/80/IPT1, L. major MHOM/SU/73/5-ASKH, L. donovani MHOM/IN/80/DD8 and L. tropica MHOM/SU/74/K27). One hundred and twenty-nine skin lesions, spotted on filter paper, were collected from patients consulting for suspicion of cutaneous leishmaniasis (CL) at the parasitology laboratory of Constantine Hospital (Algeria). Ninety-seven (75.2%) of the samples analyzed were positive. Sixty-one (5%) were related to L. major strain. These results indicate that PCR assay provides pleasant results with filter paper and represents a tool for the identification of old world CL.
