RESUMO
Abstract Listeria monocytogenes is responsible for causing listeriosis, a type of food poisoning with high mortality. This bacterium is mainly transmitted to humans through the consumption of contaminated foods. Detection of L. monocytogenes through molecular methods is crucial for food safety and clinical diagnosis. Present techniques are characterized by low discrimination power and high cost, as well as being time-consuming and taking several days to give the final result. In our study, MLVA-HRM (Multiple-Locus Variable-number tandem repeats Analysis ‒ High-Resolution Melting) was investigated as an alternative method for a fast and precise method for the genotyping of L. monocytogenes isolates. Forty-eight isolates of L. monocytogenes obtained from the microbial bank of Department of Microbiology, Iran University of Medical Sciences, were typed by MLVA-HRM analysis using five Variable Numbers of Tandem Repeat (VNTR) loci. A total of 43 different types were obtained. This research demonstrated the usefulness of the MLVA-HRMA method and its ability to discriminate L. monocytogenes isolates. Since this method is easier and more efficient than existing methods, it can be widely used in food processing plants and diagnostic laboratories as a fast and accurate method.
RESUMO
Acinetobacter baumannii is one of the most important etiologies of nosocomial infections in recent years mainly because of increasing in frequency of multidrug and pan-resistant pathogens. Meningitis caused by this organism is a dilemma; because polymyxins are the only effective antibiotics against pan-resistant serotypes, but have poor penetration via blood brain barrier; however, it has still remained uncertain whether the intravenous therapy with these agents is an effective treatment with the sufficient concentration of the drug in the cerebrospinal fluid. Herein, we report a neonate who suffered from pan-resistant A. baumannii nosocomial meningitis successfully treated with intravenous colistin combined with meropenem and rifampin. It seems that intravenous colistin at least in combination with rifampin and meropenem might be considered as an option to try in patients in whom daily intrathecal injection or insertion of intraventricular device is not possible.
RESUMO
Acinetobacter baumannii is one of the most important etiologies of nosocomial infections in recent years mainly because of increasing in frequency of multidrug and pan-resistant pathogens. Meningitis caused by this organism is a dilemma; because polymyxins are the only effective antibiotics against pan-resistant serotypes, but have poor penetration via blood brain barrier; however, it has still remained uncertain whether the intravenous therapy with these agents is an effective treatment with the sufficient concentration of the drug in the cerebrospinal fluid. Herein, we report a neonate who suffered from pan-resistant A. baumannii nosocomial meningitis successfully treated with intravenous colistin combined with meropenem and rifampin. It seems that intravenous colistin at least in combination with rifampin and meropenem might be considered as an option to try in patients in whom daily intrathecal injection or insertion of intraventricular device is not possible.
RESUMO
Lipopolysaccharide [LPS] is an important structural component of the outer cell membrane complex of gram negative microorganisms. Its causative role in gram negative bacteria-induced diseases and broad applications in different kinds of cell stimulation experiments provided a conceptual basis for studies directed at the isolation, purification, and detailed chemical characterization of LPS. The main problem with LPS purification protocols is the contamination of the end product with nucleic acids and proteins in variable proportions which could potentially interfere with downstream applications. In this study, a simple procedure for purification of LPS from Escherichia coli [E.coli] and Salmonella typhi [S.typhi] with high purity and very low contaminating nucleic acids and proteins based on the hot phenol-water extraction protocol has been introduced. The purity of extracted LPS was evaluated by silver and coomassie blue staining of SDS-PAGE gels and HPLC analysis. Limulus Amebocyte Lysate [LAL] coagulation activity and rabbit pyrogen assay were exploited to monitor the functionality of purified LPS. The results showed that DNase and RNase treatment of the sample is essential after the sonication step to eliminate nucleic acid contamination in the LPS fraction. Silver staining demonstrated ladder pattern which is characteristic of LPS. No contaminating protein was found as assessed by coomassie blue staining. HPLC fractionation revealed high degree of purity comparable with commercial LPS. Parenteral administration of purified LPS resulted in substantial increase of rabbits' body temperature [mean: 1.45°C]. LAL coagulation assay confirmed the functional activity of the purified LPS. In conclusion, the protocol presented here could be employed for isolation of LPS with high purity and functional activity