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Biomedical subjects

V Ritacco

Publications and source records attributed to V Ritacco.

31 records · Page 2Linked to original sources
PubMed1991

[Usefulness of the enzyme immunoassay in the diagnosis of tuberculosis].

To assess the usefulness of enzyme immunoassay as a rapid method of diagnosing tuberculosis, a study was conducted of 687 serum samples from 271 children and 416 adults. With 55 sera from nontuberculous children as controls, the specificity was 0.98, and with 137 controls from the adult population, 0.93. Prior vaccination with BCG did not influence the level of detectable anti-PPD antibody. The results were similar in healthy PPD-positive and negative adults. The test differentiated mycoses and nontuberculous mycobacterioses from tuberculosis. The sensitivity rates in 49 children and 200 adults diagnosed with bacteriologically confirmed pulmonary and extrapulmonary tuberculosis were calculated at 0.51 and 0.69, respectively. In those tuberculosis cases not bacteriologically confirmed or at other sites, the test was positive in 28.1% of 114 children and in 48.6% of 35 adults. The cost, speed, and availability of reagents for this test were comparable to those for direct microscopic examination. Both methods were positive for 49% of the tuberculosis cases confirmed by culture, and a total of 84% of those cases were found positive using one method or the other. It is concluded that enzyme immunoassay can be especially useful in the rapid diagnosis of nonbacilliferous pulmonary, extrapulmonary, and childhood tuberculosis.

Adolescent↗
PubMed1990

Further evaluation of an indirect enzyme-linked immunosorbent assay for the diagnosis of bovine tuberculosis.

The sensitivity and specificity of an ELISA for the detection of bovine IgG anti-Mycobacterium bovis antibodies were 73.6% and 94.1%, respectively, as determined in 53 bacteriologically confirmed tuberculous cattle and 101 healthy cattle from a tuberculosis-free area. In addition, the results of ELISA and tuberculin tests in 149 cattle were compared with those of subsequent necropsy studies. Both tests failed to detect 2 animals with tuberculous lesions and positive culture; 3/12 cattle with M. bovis isolation and no lesions, and 2/7 with atypical mycobacterial infection reacted to tuberculin, but none had antibodies; in 128 cattle with neither lesions nor mycobacterial isolation, 6 were tuberculin reactors and 7 others had antibodies. Negative results were obtained by ELISA in 21/22 paratuberculous cattle. Antibodies were not detected in 88.9% to 96.4% of 697 cattle from two tuberculin negative herds of an endemic area. In a herd with proved M. bovis infection, distribution of seropositive animals in tuberculin and non-tuberculin reactors was similar. Antibody responses to cutaneous tuberculin stimuli were observed in 4 experimentally infected cattle, but only in 2/10 healthy controls after repeated PPD stimuli. Nine controls which had either received a single tuberculin dose or none showed no increase in antibody levels. The low sensitivity of this ELISA limits its usefulness as a diagnostic tool for bovine tuberculosis eradication campaigns. However, it could be helpful in epidemiological surveillance if its efficiency to identify infected herds is demonstrated.

Animals↗
PubMed1989

Detection of circulating antibodies to purified protein derivative by enzyme-linked immunosorbent assay: its potential for the rapid diagnosis of tuberculosis.

An enzyme-linked immunosorbent assay for detecting antibodies to purified protein derivative was evaluated as a rapid method for serodiagnosis of childhood tuberculosis. Its specificity for IgG antibodies was 0.98 as determined in 55 sera from nontuberculous children who showed no significant effect of previous Bacillus Calmette-Guérin vaccination on the production of specific antibodies. Results were negative in 29 of 33 (87.9%) tuberculin-positive children and in 18 of 20 (90.0%) contacts, none of whom had evidence of tuberculosis. The sensitivity of this test was 0.51 as determined in 49 sera from bacteriologically confirmed cases; 17 of 27 smear positive cases and 8 of 22 children with positive cultures were detected. Results were positive in 32 of 114 (28.1%) patients with a diagnosis of tuberculosis not confirmed by microbiology. Consequently whereas a negative result does not rule out tuberculosis, a positive result is a strong indication of the disease. The IgM antibody determination yielded much less discriminative results.

Adolescent↗
PubMed1989

[Evaluation of an enzyme immunoassay for the rapid diagnosis of paucibacillary tuberculosis in adults].

The ELISA has been extensively evaluated as a serodiagnostic method for tuberculosis. However, there is scarce information about its application to cases that cannot be diagnosed by microscopic examination: those with closed lesions or undergoing early stages of the disease. Since a reliable serological test might substantially contribute to their prompt detection, the objective of the present study was to determine the diagnostic value of an ELISA applied to adult smear-negative cases of tuberculosis. Sera from 235 patients with active tuberculosis--176 pulmonary and 59 extrapulmonary cases--and 181 control subjects were tested for IgG antibodies to PPD by ELISA. Eleven cases of non tuberculous mycobacterial (MOTT) disease and 33 cases of mycosis were also included in this group. With the adopted cut-off value, 73.9% (105/142) of smear positive and 52.7% (49/93) of smear negative tuberculosis cases, were correctly classified. Particularly in the latter, the test was positive in 55.2% (32/58) of patients with positive cultures for Mycobacterium tuberculosis and in 48.6% (17/35) of patients diagnosed by clinical, radiological and or histopathological findings. No antibody activity was demonstrated in 92.7% of sera from the control population which included 92 healthy volunteers, 32 non tuberculous diseased subjects and 13 household contacts of smear-positive cases. Among those control subjects who were skin tested, ELISA results were not related to the tuberculin reactivity: 93.7% (30/32) of tuberculin negative and 95.2% (40/42) of tuberculin positive healthy individuals had no detectable antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1979

Immunofluorescent anti-Junin virus antibodies in Argentine hemorrhagic fever.

Immunofluorescent anti-Junin virus antibodies were detected in 15 nonfatal cases of Argentine hemorrhagic fever between the 2nd and 3rd week after onset of symptoms. In most cases, antibodies appeared by the day of clinical improvement, or between 2 and 4 days later. It was interesting to note that in 5 of 11 cases studied, the first positive serum sample presented anti-Junin virus immunofluorescent antibodies in IgA. In 2 of these 5 cases, IgA was the only immunoglobulin with antibody activity in the early positive serum sample.

Animals↗
PubMed1979

Circulating antibodies to peripheral nerve in American trypanosomiasis (Chagas' disease).

An antibody reacting with Schwann sheaths of myelinated somatic and unmyelinated autonomic peripheral nerve was found in sixty-one out of seventy-one chronic, and nine out of ten acute, Chagas' disease sera. Indirect immunofluorescence (IFL) was carried out on rat, mouse and human somatic nerves and rat sympathetic nerves with initial serum dilutions of 1 : 10, and the staining reached a final titre of 1 : 320 in some cases. The antibodies fixed complement and were absorbed out by lyophilized epimastigotes of T. cruzi. Lipid extraction of the tissue sections enhanced the staining of myelinated nerve, whereas unfixed unmyelinated sympathetic nerve was strongly reactive. Central nervous tissue did not display any positive staining on neurons, glial cells or periaxonal sheaths. Furthermore, by using a double-labelled IFL technique, it was possible to show that a rabbit antiserum raised against guinea-pig spinal cord and the chagasic anti-nerve antibodies reacted with different structures in the rat sciatic nerve. These findings suggest that the reactive antigen(s) could be located on Schwann cells. The majority, but not all, of the chagasic individuals with anti-nerve antibodies also showed the sarcolemmal and endothelial staining (EVI) previously described in Chagas' disease. The possible recognition of Schwann cell antigens by circulating antibodies in Chagas' disease could be relevant, since an autonomic denervation has been postulated as a pathogenic mechanism of cardiomyopathy and megaviscera in this condition.

Animals↗
PubMed1975

Ultrastructural and immunohistochemical studies in five cases of Argentine hemorrhagic fever.

Ultrastructural and immunohistochemical studies on tissues from five patients with Argentine hemorrhagic fever revealed previously undetected lesions caused by the viral infection. Two types of particle were seen in the cells of all organs examined. The particles had some characteristics similar to those described for arenaviruses. However, the virus-like particles were intracellular, had a single membrane, and apparently originated by a process of budding into the endoplasmic reticulum cisternae. Intranuclear bodies and three types of cytopolasmic change were observed in conjunction with the virus-like particles; Antigenic determinants of Junin virus were demonstrated in cells of all organs examined. Immunohistochemical experiments also indicated alterations in the cellular mechanisms of protein synthesis. Until now the pathogenesis of human diseases produced by arenaviruses has not been established. The results of this study suggest that in Argentine hemorrhagic fever the virus is responsible for a direct pathogenic action.

Adult↗
PubMedDate not supplied

[Study of Mycobacterium bovis in milk using bacteriological methods and the polymerase chain reaction].

The frequency of Mycobacterium bovis detection in milk samples obtained from infected animals was explored in an intensive dairy area in Argentina. To this end, an "in house" polymerase chain reaction (PCR) method was developed using Mycobacterium tuberculosis Complex specific INS1-INS2 primers, and its performance was compared with that of bacteriological methods. The decontamination procedures previous to culture reduced M. bovis viability. The pathogen was identified in milk samples from 1 of 143 infected cows and in none of 43 uninfected ones. Even though PCR sensitivity was found to be 2-20 times higher than that of bacteriology in experimentally inoculated milk samples, all 186 field samples resulted negative by PCR, including the bacteriologically-confirmed one. In spite of the high prevalence of bovine tuberculosis in Argentinian dairy herds, the detection of M. bovis in milk is an unusual finding.

Animals↗
PubMedDate not supplied

[False-positive cultures due to cross contamination in tuberculosis laboratories].

Fifteen episodes of Mycobacterium tuberculosis laboratory cross-contamination suspected between 1996 and 2001 at 6 laboratories in Buenos Aires City and suburbs were investigated by IS6110 RFLP. Thirteen episodes were confirmed. Even though BACTEC 460 produced the highest number of confirmed episodes in a single laboratory, the most extended one occurred while employing conventional culture procedures in solid medium. The double repetitive element-polymerase chain reaction (DRE-PCR) was applied to 8 of these episodes and produced concordant results with those of the RFLP. The DRE-PCR appears to be a valuable tool for the prompt identification of false positive cultures. The timely rectification of defects in laboratory protocols can avert false diagnoses of tuberculosis and unnecessary prolonged treatments.

Aerosols↗
PubMedDate not supplied

Comparative molecular study of Mycobacterium tuberculosis strains, in times of antimicrobial drug resistance.

Strains of Mycobacterium tuberculosis were compared using two DNA fingerprinting techniques: Restriction Fragment Length Polymorphism (RFLP) and Double-Repetitive-Element-PCR (DRE-PCR). Two of these strains: IH1 (susceptible to isoniazid) and IH2 (resistant to isoniazid) were recovered from cases of pulmonary tuberculosis which occurred in two brothers who lived together. The first one was recognized on July 1999, and the second was diagnosed one year later. IH1 and IH2 showed the same pattern of bands with both molecular tests. These results suggest that single drug chemoprophylaxis may occasionally select resistant strains for that drug, which can eventually cause disease and be recognized through these tests. Strains IH3, IH4 and IH5 were obtained from sputum samples of 3 different patients, and intra-laboratory cross-contamination was suspected when it was realized that the 3 positive materials had been consecutively processed the same day by the same worker in the same biological safety cabinet. Again, the 3 strains revealed identical band patterns with RFLP and DRE-PCR, confirming the posed suspicion. The results with DRE-PCR were obtained after only 8 hours of work, without the need for subcultures. This procedure allows quick correction of treatment conducts, avoiding unnecessary exposure of people and bacteria to antimicrobial drugs.

Adult↗
PubMedDate not supplied

[Evaluation of four antigens for the detection of anti-Mycobacterium bovis antibodies by enzyme immunoassay].

An enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine tuberculosis through the detection of specific seric antibodies has recently been developed in our laboratory. In order to assess its reproducibility and select the most adequate antigen, four bovine PPDs from different sources were evaluated in parallel: PPD M. bovis strain AN5, CEPANZO standard (CPZ), PPD M. bovis strain AN5, European Economic Community standard (EEC), PPD M. bovis strain AN5, prepared from non heated bacilli, killed by phenol (P) and PPD. M. bovis BCG strain prepared at the Pasteur Institute, Paris (BCG). Sera from 22 healthy cattle from tuberculosis free area and 20 bacteriologically confirmed tuberculous animals were employed in simultaneous assays. Antibody mean and standard deviations from healthy cattle expressed as optical density (OD) values were 45 +/- 22 when CPZ was used as antigen, 24 +/- 10 with EEC, 103 +/- 56 with P and 56 +/- 20 with BCG. Mean O.D. from tuberculous cattle were 588 +/- 158, 510 +/- 234, 782 +/- 138 and 441 +/- 189 with antigens CPZ, EEC, P and BCG respectively. A close correlation was observed when results obtained with EEC and P were compared with that of CPZ (r: 0.97 and 0.94 respectively). A lower specificity was achieved when BCG was used as antigen being also lower its correlation with the results obtained with CPZ (r: 0.87). It is concluded that our ELISA would achieve similar sensitivity and specificity if CPZ, EEC and P were used as antigens. On the other hand, BCG would not be suitable for this assay.

Antibodies, Bacterial↗