ABSTRACT
A screening test for the diagnosis of tuberculosis by immunodot (IDt) is described, using an antigen of Mycobacterium tuberculosis, namely, a 38-kDa glycoprotein which has shown great specificity in previous serologic analyses. The test was used to examine 28 sera from patients with lung tuberculosis. Of these, 85% were positive by micro-ELISA and by the IDt test herein described. Control sera from healthy subjects (n = 20) gave negative results for ELISA and for IDt, which indicates that the screening test is highly specific. The test is easy to handle and requires no equipment and is therefore particularly useful for field studies.
Subject(s)
Antibodies, Bacterial/blood , Glycoproteins/immunology , Mycobacterium tuberculosis/immunology , Tuberculosis/diagnosis , Antigens, Bacterial/immunology , Enzyme-Linked Immunosorbent Assay , Humans , Molecular Weight , Sensitivity and Specificity , Tuberculosis/microbiologyABSTRACT
The cellular distribution of 65 and 70 kD heat shock proteins (HSPs) was studied in the normal rat kidney and after acute tubular necrosis (ATN) induced by inorganic mercury (HgCl2). In the normal kidney the 65 kD HSP was found in the cytoplasm of podocytes and proximal convoluted tubules, whereas the 70 kD HSP was located in nuclei and cytoplasm of podocytes, cortical convoluted, and collecting tubules. The distribution of both HSPs along ATN changed as a function of time. In the early phase, before evidence of histological damage, both HSPs were found in the pielocaly ceal epithelium and medullary collecting tubules. During the necrotic phase, HSPs coexisted with sites of severe damage (i.e. cortical tubules). With immunoelectron microscopy damaged cells showed an abundance of 65 kD HSP-I in mitochondria, as well as in chromatin and nucleoli, while 70 kD HSP-I was overexpressed in the cytoplasm, mito chondria, lysosomes, cytoskeleton, chromatin, and nucleoli, and coincided with urinary excretion of HSPs. In the postregenerative phase, the distribution of HSPs was similar to that found in the normal kidney. HSPs of 65 and 70 kD were encountered constitutionally and their immunolabeling is correlated with the magnitude of cell injury.
Subject(s)
Chaperonins/chemistry , HSP70 Heat-Shock Proteins/chemistry , Kidney Tubular Necrosis, Acute/pathology , Kidney Tubular Necrosis, Acute/urine , Animals , Bacterial Proteins/urine , Chaperonin 60 , Chaperonins/urine , HSP70 Heat-Shock Proteins/urine , Kidney/chemistry , Kidney Tubular Necrosis, Acute/chemically induced , Male , Mercuric Chloride/toxicity , Rats , Rats, Wistar , Subcellular Fractions/chemistry , Subcellular Fractions/drug effects , Subcellular Fractions/pathologyABSTRACT
In this work, we grew Mycobacterium tuberculosis in an enriched Proskauer-Beck-Youmans culture medium in the presence and in the absence of phosphate salts. Immunoblot analysis of sonic extracts showed overexpression of the 38-kDa protein antigen by bacilli grown in the medium without phosphate. These observations were confirmed by immunogold electron microscopy, which showed that the number of gold particles was significantly higher in bacilli grown in medium without phosphate than in bacilli grown in medium with phosphate. The 38-kDa protein was located mainly in the wall and on the cell surface.
