RESUMO
SUMMARY: The appearance of Pseudomonas aeruginosa strains with multi-resistance to antibiotics is a clinical problem of great relevance. The methods for detecting these resistances are laborious and slow, which is a complication when treating patients promptly. In this work, we developed a simple method for simultaneous detection of several carbapenem resistance genes using a multiplex PCR assay. The PCR assay developed, followed by electrophoretic separation of fragments, allows to simultaneously identify the presence of 6 antibiotic resistance genes: bla-VIM (261 bp), bla-IMP (587 bp), bla-SPM (648 bp), bla-GIM-1 (753 bp), bla-NDM-1 (813 bp) and bla-KPC (882 bp). We analyzed 7 clinical isolates of P. aeruginosa obtained in Chile, finding the resistance genes bla-VIM, bla-IMP, bla-SPM, bla-GIM, and bla-NDM in 5 of them. We found a perfect correlation between the detection of various resistance genes by PCR and the results obtained by antibiograms. Interestingly, 2 of the strains possessed 3 different resistance genes simultaneously. Finally, in this work, we found the presence of 3 genes never described before in clinical isolates of P. aeruginosa in Chile (bla-IMP, bla-SPM, and bla-GIM-1). We developed a rapid multiplex PCR test for the simultaneous detection of up to 6 antibiotic resistance genes of the metallo-β-lactamase family in P. aeruginosa.
La aparición de cepas de Pseudomonas aeruginosa con resistencias a diversos antibióticos es un problema clínico de gran relevancia. Los métodos de detección de dichas resistencias son laboriosos y lentos, lo que genera una complicación al momento de tratar a los pacientes oportunamente. En este trabajo desarrollamos un método simple de detección simultánea de varios genes de resistencia a carbapenem, mediante un sistema de PCR múltiple. El ensayo de PCR desarrollado, seguido de una separación electroforética de los amplicones, permite distinguir simultáneamente la presencia de 6 genes de resistencia a antibióticos: bla-VIM (261 pb), bla-IMP (587 pb), bla-SPM (648 pb), bla-GIM-1 (753 pb), bla-NDM-1 (813 pb) y bla-KPC (882 pb). Analizamos 7 aislados clínicos obtenidos en Chile, encontrando en 5 de ellos los genes de resistencia bla-VIM, bla-IMP, bla-SPM, bla-GIM y bla-NDM. Encontramos una perfecta correlación entre la detección de diversos genes de resistencia y los resultados obtenidos mediante antibiogramas. Interesantemente, 2 de las cepas mostraron poseer simultáneamente 3 genes de resistencia distintos. Por último, en este trabajo encontramos la presencia de 3 genes nunca antes descritos en aislados clínicos de P. aeruginosa en Chile (bla-IMP, bla-SPM y bla-GIM-1). Hemos desarrollado un test rápido de PCR múltiple, para la detección simultánea de hasta 6 genes de resistencia a antibióticos de la familia.a de las metallo-b-lactamases en P. aeruginosa.
Assuntos
Pseudomonas aeruginosa/enzimologia , beta-Lactamases/genética , Pseudomonas aeruginosa/genética , Farmacorresistência Bacteriana , Reação em Cadeia da Polimerase MultiplexRESUMO
The nutritional status of 111 elderly men and women aget 60 to 99 years in two institutions in the Ecuadorian Andes was evaluated using dietary, anthropometric and biochemical measurement. Mean daily energy intakes of men and women were 1720 and 1650 Kcal, respectively (7,2 and 6.9 MJ), corresponding to 1.5 times the calculated basal metabolic rate (BMR). 6 por ciento of subjects had intakes less tham 1.2 x BMR and 40 por ciento less than 1.5 x BMR. Body mass index (BMI) was below 20 kg/m2 in 21 por ciento of subjects; 45 por ciento were between 20 and 24. Only one subject had a BMI over 29. Intakes of protein, calcium, iron, riboflavin and vitamin C were satisfactory, in contrast to those of zinc, vitamin A, niacin and thiamin. Using sex-and altitude-specific cutoffs for hemoglobin, anemia was observed in 11 por ciento of subjects. Nutritional status, evaluated using biochemical indicators, was satisfactory with respect to riboflavin, vitamin A, vitamin E and copper. 14 por ciento of subjects had low serum zinc levels and 17 por ciento had serum albumin values below 35 g/L. Low physical activity was associated with low energy intake and low serum albumin levels