Evaluation of a selective chromogenic medium for detecting vancomycin-resistant enterococci

ABSTRACT Rapid identification of vancomycin-resistant enterococci (VRE) can assist in choosing the appropriate treatment and preventing VRE spread. The performance of chromIDTM VRE agar was evaluated using 184 clinical isolates of Enterococcus spp. and reference strains. The test had a sensitivity of 95.52% but a low specificity of 30%.(AU)

[Optimization of screening methodologies for the detection of Streptococcus agalactiae in pregnant women]. / Optimización de metodologías de cribaje para la búsqueda de Streptococcus agalactiae en embarazadas.

Streptococcus agalactiae is a significant worldwide cause of morbidity and mortality in pregnant women and their newborn infants. The objective of this work was to determine the usefulness of bioMerieux chromogenic medium chromID Strepto B (CR) for detecting S. agalactiae in pregnant women from the selective Todd-Hewitt broth (sel-THB ) against the methods proposed by the CDC . A total of 1924 swabs were analyzed, 962 from vaginal introitus and 962 rectal, belonging to 962 women in weeks 35-37 of pregnancy. The swabs were directly seeded in CR. Both swabs were later placed in sel-THB with 15 µg/ml supplement of nalidixic acid and 10 µg/ml colistin. After 24 h of incubation, subcultures in CR medium and agar containing 5% sheep blood (SBA) were performed. The prevalence found was 17.4%. Sensitivity, specificity, positive and negative predictive values of sel-THB subcultures with CR supplement and 48 h incubation were: 98.8, 100, 100 and 99.7%, respectively. The corresponding values of direct harvest of the sample were 57.8, 100, 100, and 90%, respectively. Sensitivity of sel-THB in SBA was 85%. Sel-THB subculture performance in CR was outstanding in comparison with the method proposed by the CDC.

Optimización de metodologías de cribaje para la búsqueda de Streptococcus agalactiae en embarazadas / Optimization of screening methodologies for the detection of Streptococcus agalactiae in pregnant women

Streptococcus agalactiae es una causa importante de morbimortalidad en mujeres embarazadas y neonatos en todo el mundo. El objetivo del presente trabajo fue determinar la utilidad del medio cromogénico chromID Strepto B de bioMérieux para detectar S. agalactiae en embarazadas cuando la muestra es sembrada directamente en dicho medio o después del enriquecimiento en caldo de Todd Hewitt selectivo, opciones que se compararon con la metodología propuesta por el CDC . Se analizaron 1924 hisopados, 962 de introito vaginal y 962 rectales, correspondientes a 962 embarazadas entre la semana 35 y 37 de gestación, asistidas en distintos hospitales. Los hisopados se sembraron directamente en el medio chromID Strepto B (CR) y luego se colocaron en un caldo de Todd Hewitt selectivo, suplementado con 15 µg/ml de ácido nalidíxico y 10 µg/ml de colistina (CTH-sel). Luego de 24 h de incubación, se realizaron subcultivos en el medio CR y en agar con 5% de sangre de carnero (ASO). La prevalencia global de S. agalactiae fue de 17,4%. La sensibilidad, la especificidad y los valores predictivos positivo y negativo del subcultivo en CR del material desarrollado en el CTH -sel fueron 98,8%, 100%, 100% y 99,7% respectivamente, con una incubación de 48 h. Los valores correspondientes de la siembra directa fueron 57,8%, 100%, 100% y 90%. La sensibilidad del subcultivo en ASO del material desarrollado en el CTH -sel fue del 85%. Se destaca el excelente rendimiento del subcultivo en CR luego del enriquecimiento en caldo de Todd Hewitt selectivo en comparación con el método propuesto por el CDC.

Streptococcus agalactiae is a significant worldwide cause of morbidity and mortality in pregnant women and their newborn infants. The objective of this work was to determine the usefulness of bioMrieux chromogenic medium chromID Strepto B (CR) for detecting S. agalactiae in pregnant women from the selective Todd-Hewitt broth (sel-THB ) against the methods proposed by the CDC . A total of 1924 swabs were analyzed, 962 from vaginal introitus and 962 rectal, belonging to 962 women in weeks 35-37 of pregnancy. The swabs were directly seeded in CR. Both swabs were later placed in sel-THB with 15 µg/ml supplement of nalidixic acid and 10 µg/ml colistin. After 24 h of incubation, subcultures in CR medium and agar containing 5% sheep blood (SBA) were performed. The prevalence found was 17.4%. Sensitivity, specificity, positive and negative predictive values of sel-THB subcultures with CR supplement and 48 h incubation were: 98.8, 100, 100 and 99.7%, respectively. The corresponding values of direct harvest of the sample were 57.8, 100, 100, and 90%, respectively. Sensitivity of sel-THB in SBA was 85%. Sel-THB subculture performance in CR was outstanding in comparison with the method proposed by the CDC.

Comparison of endotoxin levels in previous studies on primary endodontic infections.

BACKGROUND: This study was performed to determine which of the quantitative methods, namely, chromogenic endpoint, chromogenic kinetic, and turbidimetric kinetic ones, best fit for the analysis of primary endodontic infections. METHODS: Twenty-one root canals with apical periodontitis were sampled with paper points. The same sample was analyzed by means of the endpoint chromogenic Limulus amebocyte lysate (LAL) assay (QCL), quantitative kinetic chromogenic LAL assay (KQCL), and kinetic turbidimetric LAL assay (Turbidimetric). RESULTS: All three LAL methods were effective in the recovery of endotoxin from root canal infection. Regardless of the method tested, endotoxin was detected in 100% of the root canals (21/21). The KQCL assay yielded a median value of endotoxin of 7.49 EU/mL, close to and not significantly different from those for the turbidimetric test (9.19 EU/mL) (both kinetic methods) (p > 0.05). In contrast, the endpoint QCL showed a median value of 34.20 EU/mL (p < 0.05). The comparison of the three methods revealed that both turbidimetric and KQCL methods were more precise, with best reproducibility (the coefficient variation between analysis of the root canal and its duplicate was lower than 10%). The inhibition/enhancement assay indicated a good interaction between the root canal samples with the turbidimetric method. CONCLUSION: This study has revealed that quantitative kinetic-turbidimetric and kinetic-chromogenic LAL methods are best fitted for the analysis of endotoxins in root canal infection, both being more precise and allowing better reproducibility compared with the endpoint-QCL assay.

Intraspecies differences in hemostatic venom activities of the South American rattlesnakes, Crotalus durissus cumanensis, as revealed by a range of protease inhibitors.

Crotalus durissus cumanensis is an endemic rattlesnake found in Venezuela and Colombia. In this study, a comparative analysis of hemorrhagic, coagulation and fibrino(geno)lytic activities in the presence or absence of protease inhibitors was performed with venoms of the same species Crotalus durissus cumanensis, from seven geographical regions of Venezuela (Lagunetica, Santa Teresa, Carrizales, Guarenas, Anzoátegui, Margarita and Maracay). Lagunetica, Carrizales and Anzoátegui venoms induced hemorrhagic activity. All venoms, except that of snakes from the Carrizales region presented thrombin-like activity, which was inhibited completely by phenylmethanesulfonyl fluoride and ethylene glycol-bis-N, N,N',N'-tetraacetic acid. This effect of the latter could be explained by the high chelant calcium effect, which is a cofactor for the fibrin polymerization process. Soybean trypsin inhibitor was effective on Santa Teresa venom. Antithrombin III/Hep complex and phenantroline partially inhibited this activity in all venoms except Margarita and Anzoátegui, respectively, which were not inhibited. Serine protease inhibitors were more effective against thrombin, kallikrein and plasmin-like amidolytic activities. Additionally, metalloprotease inhibitors significantly inhibited the t-PA-like amidolytic activity and completely the hemorrhagic and fibrino(geno)lytic activities. In conclusion, the thrombin-like activity observed in these venoms was partially reduced by serine protease inhibitors, indicating the possible presence of catalytic domains in those enzymes that do not interact with these inhibitors. Using protease inhibitors on venom hemostatic activities could contribute to our understanding of the active components of snake venom on the hemostatic system, and further reveal the intraspecies variation of venoms, which is important in the treatment of envenomation, and in addition represents an interesting model for the study of venom in hemostasis.

Assessment of chromogen suitability in ELISA for the detection of anaplasmosis and trypanosomosis.

Two different ELISAs were routinely performed in our laboratory to detect bovine trypanosomosis and anaplasmosis. The ELISA test for trypanosomosis involved the adsorption of a soluble fraction of parasites as the antigen; and, the ELISA for anaplasmosis was performed with a purified recombinant protein MSP5r adsorbed to the plate. With the purpose of assessing the merit of ABTS and TMB, we compared the absorbance obtained from positive and negative control sera from both assays. The results obtained, suggest that TMB is more adequate for recombinant antigens and that ABTS is preferred when partially purified antigenic extracts are used in the ELISA test.

Método para cuantificar actividad proteolítica sobre sustratos insolubles coloreados / Method for quantitative proteolytic activity about stained substrates insoluble

Se cuantificó la actividad proteolítica de Staphylococcus aureus aislados de infecciones del hombre, aplicando una modificación de la técnica efectuada por otros autores con Pseudomonas aeruginosa. Además del polvo de piel coloreado con Azul Brillante de Remazol utilizado con esta bacteria, se prepararon otros sustratos insolubles, tales como elastina-Rojo Congo, colágeno-Rojo Congo y polvo de ubre-Rojo Congo. Fue posible comprobar mayor actividad proteolítica sobre colágeno que sobre elastina, mientras que con los extractos o polvos tisulares se evidenció más proteólisis con polvo de ubre de vaca (PUH) que con polvo de piel (PPA). Cabe destacar que las cepas procedían de infecciones humanas, incluyendo afecciones no epidérmicas y una de colección ATCC. El método desarrollado constituyó una ventaja con respecto a las técnicas clásicas de bacteriología que detectan cualitativamente la actividad proteolítica

Método para cuantificar actividad proteolítica sobre sustratos insolubles coloreados / Method for quantitative proteolytic activity about stained substrates insoluble

Se cuantificó la actividad proteolítica de Staphylococcus aureus aislados de infecciones del hombre, aplicando una modificación de la técnica efectuada por otros autores con Pseudomonas aeruginosa. Además del polvo de piel coloreado con Azul Brillante de Remazol utilizado con esta bacteria, se prepararon otros sustratos insolubles, tales como elastina-Rojo Congo, colágeno-Rojo Congo y polvo de ubre-Rojo Congo. Fue posible comprobar mayor actividad proteolítica sobre colágeno que sobre elastina, mientras que con los extractos o polvos tisulares se evidenció más proteólisis con polvo de ubre de vaca (PUH) que con polvo de piel (PPA). Cabe destacar que las cepas procedían de infecciones humanas, incluyendo afecciones no epidérmicas y una de colección ATCC. El método desarrollado constituyó una ventaja con respecto a las técnicas clásicas de bacteriología que detectan cualitativamente la actividad proteolítica