Bacteriology of community acquired urinary tract infection. Analysis of 1048 cases.

Urine samples from 1048 cases clinically suspected to have urinary tract infection were examined for bacteriological culture and antibiotic sensitivity. Of 1048 cultures, 530 (50.5%) were sterile, 163 (15.5%) showed the presence of nonpathogenic bacteria and 11 (1%) revealed nonsignificant bacteriuria with Escherichia coli. Significant bacteriuria (more than 10(5)/ml) was found in the remaining 344 (32.8%) cases. Escherichia coli was the predominant pathogen isolated from 170 (16.2%) samples; Klebsiella aerogenes was the next in order, found in 46 (4.3%) cases. Streptococcus pyogenes was isolated in 37 (3.5%), Pseudomonas aeruginosa in 29 (2.7%), Proteus mirabilis in 26 (2.4%), Streptococcus faecalis in 21 (2.0%), Staphylococcus aureus in 12 (1%) and Citrobacter freundii in 3 cases. Nalidixic acid was the most effective drug against gram negative organisms. Nitrofurantoin was the next antibiotic of choice, whereas cephalexin was the third drug in order of sensitivity. Ampicillin was the first drug of choice in infection with gram positive cocci, followed by nitrofurantoin, tetracycline, cephalexin and cephaloridine. The results of sensitivity tests emphasise the problem of multiple drug resistance in our community.

Raising monospecific antibodies by use of protein components prestained with Remazol Brilliant Blue and separated by disc electrophoresis on polyacrylamide gel.

We have reported (Clin Chem 29: 42-44, 1983) that prestaining with Remazol Brilliant Blue permits direct visualization of serum components on disc electrophoresis, and apparently purifies the proteins well. Here we have cut out the bands corresponding to the prestained albumin and transferrin after disc electrophoresis of normal human serum proteins, eluted some individual proteins into saline, and assessed their purity by immunoelectrophoresis and two-dimensional crossed immunoelectrophoresis against polyvalent antihuman serum. These two techniques indicated purity of these antigens. We inoculated rabbits with the eluates containing the pure antigens, and tested the resulting antibodies for monospecificity by immunoelectrophoresis, rocket electrophoresis, and single radial immunodiffusion. From the results we conclude that the antibodies raised against each component were monospecific, and that this is a simple, economical, rapid, and reliable method for obtaining a pure fraction of serum protein for use as an antigen.

Remazol Brilliant Blue as a pre-stain for the immediate visualization of human serum proteins on polyacrylamide gel disc electrophoresis.

We describe the pre-staining of proteins in normal human serum with Remazol Brilliant Blue before separation by disc electrophoresis. Critical to reproducibility are: dye concentration of 0.16 mol/L in a Tris-glycine buffer (pH 8.3), use of equal volumes of serum and dye solution, a tagging period of 2 h at room temperature, and electrophoresis of 0.1 mL of the mixture at 2.5 mA/gel for about 2 h. Advantages include speed, quality of resolution, and low background. This method was compared with Amido Black post-electrophoresis staining in 35 sera. Of these, 16 showed identical results with respect to the number of bands. In the other 19, Amido Black showed more bands in the post-transferrin region and, sometimes, in the post-albumin region. The pre-stained gels showed slower electrophoretic mobilities of the components. Protein bands eluted from pre-stained gels retained immunological reactivity.

A Comparison of Antigens of Trichophytion Rubrum Obtained by Some Methods of Preparation.

Westphal's and Reyes and Friedman's methods of extraction were used to study the antigens of Trichophyton rubrum. Three types of antigens were compared - phenolic (P) and aqueous (A) materials from mechanically ground organisms, precipitated by increasing gradients of ethanol and extra cellular (X) metabolites (concentrated medium) precipitated by increasing gradients of acetone. There was no yield after concentration of 75% for both ethanol and acetone. The extra cellar material (X) had the maximum nitrogen content. Polyacrylamide gel disc electrophoresis separations stained by PAS and amido black showed a few common PAS positive bands in the three polyvalent extracts. (X) was the richest in protein and showed four bands. Antisera -,zre raised in rabbi using (P) , (A) and (X) material. (X) was an effective antigen showing two immuno - precipitates with homologous antibody and one each with (P) and (A) in agar gel double diff usion (AGD) and as many as eight components in two dimensional crossed immunoelectrophoresis (2D - CIEP). (A) and (P) materials were poorly antigenic. Antisera to these had no reactivity in AGD and showed only one and two precipitinogens respectively in 2D - CIEP.