Introduction: Methicillin resistant Staphylococcus aureus (MRSA) causes illness to people and can be picked up from both healthcare facilities and the environment leading to high morbidity and mortality. The study was aimed at identifying phenotypic characteristics of Methicillin-Resistant Staphylococcus aureus and determine the antibiotic susceptibility pattern of clinical samples isolated from patients attending or admitted in two health facilities in Kiambu County, Kenya. Methodology: One hundred and thirty-eight (138) clinical samples were collected from patients attending Thika and Kiambu Level-5 Hospitals. The isolates were obtained using standard bacteriological techniques. Methicillin resistance of Staphylococcus aureus was determined using the cefoxitin disk diffusion test. Results: Out of 138 samples, 54 (39.1%) were found to have Staphylococcus aureus of which 22 (40.7%) were shown to be MRSA using the cefoxitin- based susceptibility test. Antibiotic susceptibility testing using Kirby-Bauer technique was performed on all 54 isolates. The highest sensitivity was found in chloramphenicol 46 (85.2%) and lowest in penicillin-G 8 (14.8%). Multi-Drug Resistance (MDR) was reported in 35 (64.8%) of the 54 isolates of Staphylococcus aureus. All 22 MRSA strains were found to be MDR. Conclusions: the data obtained revealed that there is presence of MRSA in healthcare settings in Kiambu County, Kenya with varying antibiotic sensitivity patterns as well as multidrug resistance. The findings will help healthcare workers in the county to develop preventive strategy as well as institute policy for antibiotic usage, infection control and surveillance.
Panton–Valentine leukocidin gene is produced by Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus isolates as a pore-forming toxin is largely responsible for skin and soft tissue illnesses. MRSA produces PVL toxins through lukS and lukF proteins causing tissue necrosis by damaging membrane of the defense cells. Presence of PVL toxin was tested from the 54 S. aureus clinical isolates obtained from Thika and Kiambu Level 5 Hospitals, in Kiambu County, Kenya, by Geno Type® MRSA assay (Hain Life Science, Nehren, Germany). DNA was isolated from freshly harvested bacterial cultures by spin column using Geno Type DNA isolation kit. The detection of PVL toxins was performed by amplification of genomic DNA and by reverse hybridization that identifies PVL genes using Geno Type MRSA kit. Out of 138 samples that were collected from patients in Kiambu County, 54 S. aureus isolates were obtained, of which 14 (25.9%; 95% CI = 11.9–38.9) samples had PVL toxins. The isolates that were obtained from the female patients had a higher PVL toxin prevalence of 35.7%, while the isolates collected from the male patients had a lower prevalence of 15.4% (P=0.09). The pediatrics department had the highest PVL gene prevalence compared to outpatient department and surgical units (P=0.08). However, the age groups of patients and the hospital attended by patients showed no significant difference in terms of PVL gene prevalence (P=0.26). Therefore, the patients' gender and hospital units were not significantly associated with PVL gene prevalence (P=0.08). This study shows that PVL positive isolates occur in the sampled hospitals in the county and female as well as children must be taken into consideration among patients with wound infections when isolating S. aureus.
This article describes the development of a new paradigm in lipid testing wherein seven major human plasma lipids are determined simultaneously without the need for analytical separations. Included are cholesterol and the esters of linoleic, conjugated linoleic (CLA), arachidonic, linolenic, eicosapentaenoic, (EPA), and docosohexaenoic acids, (DHA). The simple quantitative colorimetric assay is rapid, rugged, inexpensive and specific to the -CH=CH-CH 2 -functional group in both cyclic and acyclic structures. The critical moiety in the functional group is the a-methylene (allylic) group that triggers the reaction. Without it, a double or triple bond is not reactive. The visible spectrum for a typical plasma sample turned out to be the linear sum of the weighted contributions from all seven analytes that -given the heterogeneity of blood samples -leads to a broad diversity in the spectral patterns. To resolve the spectral data, a number of chemometric techniques were investigated. Initially, calculations were made on spectral data that originated from prepared synthetic mixtures whose molar concentrations were known. The most encouraging algorithms employed are: coupling ridge regression (RR) with K-matrix, partial least squares regression (PLS), and generalized standard addition method (GSAM). An attempt to apply GSAM to serum samples is validated using GC-MS. The same serum samples are analyzed using GC-MS and spectrophotometry and the ratios of omega-6 to omega-3 obtained from the analyses are compared. Results using both methods correspond very well. (The reaction between acetyl chloride and olefinic systems is described as a Friedel-Crafts reaction but the mechanism of this process, the structure of the products, and the nature of the resulting chromophore remains to be fully identified.)
Current methodologies of quantifying cholesterol and monounsaturated fatty acid (MUFA)/ polyunsaturated fatty acids (PUFAs) constitute techniques such as GC-MS and HPLC, which use expensive standards, are time consuming and require skilled labor. We sought to develop a simple, direct alternative method for the simultaneous determination of cholesterol and MUFA/PUFAs (linoleic (LA), alpha-linolenic (ALA), arachidonic (AA), eicosapentaenoic (EPA), docosahexaenoic (DHA), conjugated linoleic (CLA), and oleic (OA) acids) using a novel colorimetric test, the "Purdie Assay." The data were analyzed using a coded chemometric software which involves clustering algorithms, and genetic algorithm partial least squares (GAPLS). The colorimetric test with GAPLS simultaneously quantified these lipids without any separation of the analytes in human serum and vegetable oils and foods. We performed pattern recognition of biological and food samples using principal component analysis (PCA) and hierarchical clustering (HC). The assay successfully discriminated 11 clusters corresponding to different food and biological samples and also discriminated synthetic vegetable oil samples using PCA and HC corresponding to levels of prepared lipids. This study shows the wide range of possible applications of the assay as a novel, fast, and efficient tool for lipid quantification and classification.Practical applications: The novel assay coupled with GAPLS, PCA, and HC can provide an efficient tool for the direct determination and discrimination of unsaturated lipids in biological samples.
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