[Dot blot for the diagnosis of porcine cysticercosis]. / Dot blot para el diagnóstico de la cisticercosis porcina.

The Taenia solium larva causes cysticercosis in pigs, as well as economic losses for farmers and taeniosis in humans, constituting a public health problem. Infested pigs must therefore be identified before they enter the food chain. To this end, a dot blot assay was developed for the immunodiagnosis of porcine cysticercosis. A study was made of 44 pigs from different areas of Colombia that had all tested positive to cysticercosis, both by necropsy and the Western blot technique. Another group was formed comprising 44 pigs that had all tested negative to Western blot and necropsy. After analysing these 88 samples to validate the diagnostic assay, the result was a sensitivity of 86.4% and a specificity of 93.2%. The dot blot assay proved useful in diagnosing porcine cysticercosis. As the assay is easy to use in laboratories in endemic areas, as well as under field conditions, it is also appropriate for epidemiological studies.

The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease

Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70 percent of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis - a simple, economical and rapid method - was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.

The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease.

Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis--a simple, economical and rapid method--was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.

[Prevalence of Taenia solium antibodies in humans and in pigs in an endemic area of Colombia]. / Prevalencia de anticuerpos para Taenia solium en humanos y cerdos en una zona endémica colombiana.

INTRODUCTION: Cysticercosis (CC) caused by Taenia solium in humans and in pigs is endemic in many rural communities in developing countries. The use of Western blot assays (WB) to determine T. solium antibodies has become the best serological tool available to date for identifying positive individuals in field conditions. AIMS: The aim of this study was to determine the prevalence of T. solium antibodies in humans and in pigs in two rural communities in Antioquia, Colombia. PATIENTS, MATERIALS AND METHODS: Serological identification of humans and pigs with T. solium antibodies was performed using WB assays in two communities in Ituango, Antioquia. During the study, demographic variables, housing and health conditions were taken into account. Contingency tables were drawn up using c2 to compare the proportion of seronegative individuals and seropositive individuals with headache, fainting or convulsions. RESULTS: The prevalence of human and porcine CC obtained was 2.23 and 6.82% in Pascuita and 1.17 and 2.33% in Guacharaquero, both respectively. Of the 11 WB positive patients evaluated by imaging techniques, two individuals were found to have single calcifications in the TAC scan and RMI showed another to have an unspecified lesion. The prevalence of infection in humans and in pigs in two rural communities in the north of the district of Antioquia, Colombia, shows that CC is endemic and that steps must be taken to control it.

Prevalencia de anticuerpos para Taenia solium en humanos y cerdos en una zona endémica colombiana / Prevalence of Taenia solium antibodies in humans and in pigs in an endemic area of Colombia

Introducción. La cisticercosis por Taenia solium en humanos y cerdos es endémica en muchas comunidades rurales de los países en desarrollo. El uso de la inmunoelectrotransferencia (EITB) -para la determinación de anticuerpos contra T.solium-se ha constituido en la mejor herramienta serológica disponible hasta el momento para la identificación de individuos positivos en condiciones de campo. Objetivo. Determinar la prevalencia de anticuerpos para T. solium en humanos y cerdos en dos comunidades rurales de Antioquia, Colombia. Pacientes, materiales y métodos. Mediante la aplicación de la prueba del EITB, se realizó la identificación serológica de humanos y cerdos con anticuerpos para T. solium en dos comunidades de Ituango, Antioquia. Se tuvieron en cuenta variables demográficas, características de las viviendas y condiciones sanitarias. Se hicieron tablas de contingencia usando la prueba de 2 para comparar la proporción de individuos seronegativos e individuos seropositivos con cefalea, desmayos o convulsiones. Resultados. Se obtuvo una prevalencia para cisticercosis humana y porcina, respectivamente, del 2,23 y 6,82 por ciento en Pascuita y de 1,17 y 2,33 por ciento en Guacharaquero. De 11 pacientes con EITB positivo evaluados mediante imágenes, se encontraron dos individuos con calcificaciones únicas en la tomografía computarizada y uno con una lesión inespecífica en la resonancia magnética. La prevalencia de la infección en humanos y en cerdos en dos comunidades rurales al norte del departamento de Antioquia, Colombia, demuestra que la cisticercosis es endémica y que se necesitan emprender acciones que permitan su control (AU)

Introduction. Cysticercosis (CC) caused by Taenia solium in humans and in pigs is endemic in many rural communities in developing countries. The use of Western blot assays (WB) to determine T. solium antibodies has become the best serological tool available to date for identifying positive individuals in field conditions. Aims. The aim of this study was to determine the prevalence of T. solium antibodies in humans and in pigs in two rural communities in Antioquia, Colombia. Patients, materials and methods. Serological identification of humans and pigs with T. solium antibodies was performed using WB assays in two communities in Ituango, Antioquia. During the study, demographic variables, housing and health conditions were taken into account. Contingency tables were drawn up using χ2 to compare the proportion of seronegative individuals and seropositive individuals with headache, fainting or convulsions. Results. The prevalence of human and porcine CC obtained was 2.23 and 6.82% in Pascuita and 1.17 and 2.33% in Guacharaquero, both respectively. Of the 11 WB-positive patients evaluated by imaging techniques, two individuals were found to have single calcifications in the TAC scan and RMI showed another to have an unspecified lesion. The prevalence of infection in humans and in pigs in two rural communities in the north of the district of Antioquia, Colombia, shows that CC is endemic and that steps must be taken to control it (AU)

Development of a chronic chromoblastomycosis model in immunocompetent mice.

An experimental model to study chromoblastomycosis caused by Fonsecaea pedrosoi was developed in immunocompetent BALB/c mice. Unlike previous models of chromoblastomycosis, in this model a chronic, progressive infection mimicking the infection in humans was developed. This model may be suitable for use in experimental studies of chromoblastomycosis.

Evaluation of three Mycobacterium leprae monoclonal antibodies in mucus and lymph samples from Ziehl-Neelsen stain negative leprosy patients and their household contacts in an Indian community.

Mucus and lymph smears collected from leprosy patients (9) and their household contacts (44) in the Caño Mochuelo Indian Reservation, Casanare, Colombia, were examined with monoclonal antibodies (MoAb) against Mycobacterium leprae. The individuals studied were: 5 borderline leprosy (BB) patients, 4 with a lepromatous leprosy (LL), all of whom were undergoing epidemiological surveillance after treatment and 44 household contacts: 21 of the LL and 23 contacts of the BB patients. The MoAb were reactive with the following M. leprae antigens: 65 kd heat shock protein, A6; soluble antigen G7 and complete antigen, E11. All the samples were tested with each of the MoAb using the avidin-biotin-peroxidase technique and 3,3 diaminobenzidine as chromogen. The patients and household contacts studied were all recorded as Ziehl-Neelsen stain negative. The MoAb which showed optimal reaction was G7, this MoAb permitted good visualization of the bacilli. Five patients with BB diagnosis and one with LL were positive for G7; of the BB patients' household contacts, 9 were positive for G7; 7 of the LL patients' household contacts were positive for the same MoAb. MoAb G7 allowed the detection of bacillar Mycobacterium spp. compatible structures in both patients and household contacts. G7 permitted the visualization of the complete bacillus and could be used for early diagnosis and follow-up of the disease in patients.

Chromoblastomycosis murine model and in vitro test to evaluate the sensitivity of Fonsecaea pedrosoi to ketoconazole, itraconazole and saperconazole.

An experimental model of murine chromoblastomycosis and in vitro tests with Fonsecaea pedrosoi were used to test the sensitivity of this fungus to three different antimycotics. The experimental model was standardized in BALB/c mice inoculated intraperitoneally with a 10(6) CFU/ml suspension of a F. pedrosoi isolate. Clinical infection was evident after 5 days of inoculation. Three groups of 27 mice each were used in the experiment. One group was treated with ketoconazole (KTZ), another with itraconazole (ITZ) and the other with saperconazole (SPZ). Antimycotic therapy was continued for 21 days. The control group consisted of 40 mice which were inoculated, but not treated. Infection was documented by macroscopic and microscopic examination of affected tissue in addition to culture of tissue macerates. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) for the F. pedrosoi strain used were done. The in vitro results showed that SPZ was the most active with MIC 0.01 microgram/ml and MFC 0.1 microgram/ml, followed by ITZ. SPZ was also the most effective in vivo since 63% of the treated animals (p = 0.01) showed a curative effect after the observation period. We concluded that SPZ had the best in vitro and in vivo activity against F. pedrosoi.