RESULTS: Unique and specific primer pairs were designed to amplify the 8 genes. The specificity of the primers was confirmed by DNA sequencing of the nanoplex PCR products and BLAST analysis. The sensitivity and specificity of V-BiA-RE nanoplex PCR assay was evaluated against the conventional culture method. The analytical sensitivity of the assay was found to be 1 ng at the DNA level while the analytical specificity was evaluated with 43 reference enterococci and non-enterococcal strains and was found to be 100%. The diagnostic accuracy was determined using 159 clinical specimens, which showed that 97% of the clinical isolates belonged to E. faecalis, of which 26% showed the HLGR genotype, but none were vancomycin resistant. The presence of an internal control in the V-BiA-RE nanoplex PCR assay helped us to rule out false negative cases.
CONCLUSION: The nanoplex PCR assay is robust and can give results within 4 hours about the 8 genes that are essential for the identification of the most common Enterococcus spp. and their antibiotic sensitivity pattern. The PCR assay developed in this study can be used as an effective surveillance tool to study the prevalence of enterococci and their antibiotic resistance pattern in hospitals and farm animals.
OBJECTIVES: We present six Italian tourists who acquired tropical ulcers in tropical and subtropical countries.
MATERIALS & METHODS: Four males and two females acquired a skin ulcer during trips to Brazil, Malaysia, Fiji Islands, Zambia, Tanzania and India. In all patients, medical history, physical and dermatological examination, laboratory tests, bacteriological examinations and biopsy were carried out.
RESULTS: All patients were in good general health. All patients stated that the ulcer was caused by a trauma. No fever was reported. Neither lymphangitis nor lymphadenopathy were detected. The ulcer was located on a forearm in one patient, on a leg in two and on an ankle in three patients. All ulcers were malodorous and painful. Laboratory tests revealed mild leucocytosis and a mild increase in erythrocyte sedimentation rate and C-reactive protein. Results of bacteriological examinations revealed the presence of Fusobacterium spp. in five patients. Other bacteria were identified in all patients. Histopathological examination showed: necrosis of the epidermis and dermis; vascular dilatation; oedema in the dermis; massive infiltration with neutrophils, lymphocytes and histiocytes; and fragmented collagen bundles. No signs of vasculitis were observed. All patients were successfully treated with oral metronidazole (1 g/day for two weeks) and, according to antibiograms, with different systemic antibiotics.
CONCLUSION: To our knowledge, these are the first cases of tropical ulcers reported in Western tourists.