Early and rapid detection of the causative organism is necessary in tuberculosis. We present here an integrated and dedicated molecular biology system for tuberculosis diagnosis. One hundred and eighty-nine (189) biologic specimens from patients strongly suspected by clinical parameters of tuberculosis were studied by Ziehl-Neelsen staining, cultivation on a solid medium, and by a balanced heminested fluorometric PCR system (Orange G3TB) that preserves worker safety and produces a rather pure material free of potential inhibitors.
View Article and Find Full Text PDFAsian Pac J Trop Biomed
April 2011
Objective: To present an integrated molecular biology dedicated system for tuberculosis diagnosis.
Methods: One hundred and five sputum specimens from patients strongly suspected by clinical parameters of tuberculosis were studied by Ziehl-Neelsen staining, by cultivation on solid medium and by a balanced heminested fluorometric PCR system (Orange G3TB) that could preserve worker safety and produce a rather pure material free of potential inhibitors. DNA amplification was performed in a low cost tuberculosis termocycler-fluorometer.
The advances in genetics and molecular biology have raised new areas in medicine, such as pharmacogenomics, which tries to predict drug responses and toxicities based on the individual genetic variability, describing the so called: pharmacogenomic syndromes. Oncology would find this development extremely useful because of the severe toxicity of chemotherapy. There are a lot of genetic loci under investigation for their potential in predicting drug toxicity, but only three of them have showed clinical usefulness up to now.
View Article and Find Full Text PDFA novel tandemly repeated DNA structure of Toxoplasma gondii that meets the requirements assigned for satellital DNA was characterized. A DNA fragment of 1002 bp contains two different elements of repetitive DNA families named ABGTg7 and ABGTg8.2.
View Article and Find Full Text PDFThe Toxoplasma gondii rhoptry protein Rop2 was expressed in Escherichia coli as a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196-561). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196-561 as the antigen substrate. The analyzed sera were divided according to T.
View Article and Find Full Text PDFWe report the potential use of a specific Toxoplasma gondii DNA probe (ABGTg7). We applied a dot blot hybridization assay to blood samples for the diagnosis of cerebral toxoplasmosis (CT), acute toxoplasmic lymphadenopathy (ATL), and disseminated toxoplasmosis in transplant recipients (TRs). We studied a total of 84 individuals: 38 patients and 46 controls.
View Article and Find Full Text PDFAn unusual case of cerebral toxoplasmosis in an HIV negative 11 year old child is reported. Central nervous system disease was assessed immunohistochemically in a brain biopsy specimen with TgP8--a specific monoclonal antibody against Toxoplasma gondii antigens--thus confirming IgG and IgM serology, technetium scan findings, and clinical data. In addition, an active parasitaemia was confirmed by DNA in situ hybridisation assay in white cells using an ABGTg4 probe.
View Article and Find Full Text PDFDeoxyribonuclease activity was detected in E. granulosus protoscoleces from sheep hydatid cysts by electrophoresis in agarose gels of DNA fragments obtained after incubation of integral DNA with a protoscoleces preparation. Preliminary characterization showed that deoxyribonuclease activity was optimal at neutral-alkaline pH, magnesium ions were required, and it was able to digest different types of DNA, making random cuts.
View Article and Find Full Text PDFMem Inst Oswaldo Cruz
February 1995
A slide micro-immunoenzymatic assay (micro-SIA) to detect antibodies to non-particulate Toxoplasma gondii antigens is described. This assay allows the diagnosis of toxoplasmosis infection in about 1 hr. Twenty-four determinations can be performed per slide.
View Article and Find Full Text PDFJ Clin Microbiol
December 1992
Central nervous system toxoplasmosis is a life-threatening infection with a mortality rate of higher than 60%. An early and rapid diagnosis is important for effective treatment of the disease. A new approach for detection of cerebral toxoplasmosis is described here.
View Article and Find Full Text PDFGenomic DNA of Toxoplasma gondii was digested with the restriction endonuclease Hpa II and the resulting repetitive DNA sequences were visualized after electrophoresis on agarose gels and staining with ethidium bromide. Three repetitive DNA sequences were isolated and cloned in the plasmid pUC19. The recombinant plasmids (pTg8, pTg4 and pTg1) had inserts of 840, 440, and 180 basepairs, respectively.
View Article and Find Full Text PDFA rapid and simple technique for the purification of Toxoplasma gondii tachyzoites was developed. Highly purified parasites were obtained from the peritoneal exudates of infected mice by means of two consecutive discontinuous sucrose gradients run at low speed (10,000xg, 30 min). Parasites obtained by this method conserved its biological activity.
View Article and Find Full Text PDFA unique 2.2-kb mRNA is transcribed from the Q6 and Q8 genes of the mouse major histocompatibility complex. RNase protection experiments and DNA sequence analysis have mapped the 3' terminus to a site located 1110 bp downstream from exon 8.
View Article and Find Full Text PDFWe have studied chemiluminescence produced by neutrophils stimulated by opsonized zymosan in insulin dependent (IDD) and non insulin dependent (NIDD) diabetic patients. Chemiluminescence was evaluated as the integral and maximum peak, total time and time to maximum peak of the response curve to opsonized zymosan. These values were then compared with circulating immune complexes (CIC) and glucose levels.
View Article and Find Full Text PDFImmunogenetics
February 1987
By use of Southern blot analyses and low copy number probes, the fine structure of the Q region of the mouse major histocompatibility complex was studied in more detail. With a probe recognizing the even-numbered genes Q4, Q6, and Q8, it was evident that Q4 and/or the regions flanking Q4 are polymorphic, whereas Q6 and Q8, and their flanking regions are nonpolymorphic. Perhaps the most noteworthy finding is that at least two strain haplotypes, H-2k and H-2f, possessed extensive deletions in the Q region.
View Article and Find Full Text PDFHybridomas were constructed with spleen cells from mice immunized against Methanosarcina barkeri 227. The reaction with the resulting monoclonal antibodies identified two antigenic determinants. Determinant 8A is present in M.
View Article and Find Full Text PDFActa Physiol Pharmacol Latinoam
May 1986
Circulating immune complexes (CIC) were evaluated in 57 diabetic patients: 28 were insulin-dependent (IDD) and 29 were non insulin-dependent (NIDD) subdivided according to the presence or absence of microangiopathy. The following techniques were used: 1) binding to human red blood cells through C3b complement fraction and posterior radioimmunoassay with 125I labeled anti human IgG (HRBC RIA test); and 2) CIC precipitation with 3.5% polyethylenglycol (PEG test).
View Article and Find Full Text PDFPrevious results demonstrated that androgen-dependent rat specific epididymal proteins (SEP) were bound to spermatozoa during their maturation in the epididymis. This paper describes the purification of glycoprotein DE, which constitutes 40% of SEP, and the identification and semiquantitative determination of the sugars forming its oligosaccharide chain. Affinity chromatography on Sepharose-Concanavalin A produced a sample of D-E 95% pure in which 10.
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