Aerosol infection of rhesus macaques with Junin virus.

The purpose of our work was to determine if aerosols of Junin virus can infect rhesus macaques and if the disease is the same as that produced by virus inoculated parenterally. The 6 macaques exposed to the virus by aerosol became acutely ill during the 3rd week after exposure, and all died. Three died by day 21, while the remainder died after 1 month. Junin virus was found primarily in visceral organs of those animals dying before 21 days after infection and in the central nervous system tissues from animals dying later. Histological changes were similar to those reported in rhesus monkeys after parenteral Junin viral infection. Gastrointestinal necrosis, however, was less severe in aerosol-infected animals and the associated septicemia was not seen. High levels of alpha interferon were detected by the 3rd day in all infected macaques. Experimental Argentine hemorrhagic fever induced by aerosol infection in rhesus macaques was similar to that seen after parenteral challenge and mimicked closely the clinical syndrome observed in humans.

Prospective, double-blind, concurrent, placebo-controlled clinical trial of intravenous ribavirin therapy of hemorrhagic fever with renal syndrome.

A prospective, randomized, double-blind, concurrent, placebo-controlled clinical trial of intravenous ribavirin (loading dose of 33 mg/kg, 16 mg/kg every 6 h for 4 days, and 8 mg/kg every 8 h for 3 days) was conducted in 242 patients with serologically confirmed hemorrhagic fever with renal syndrome (HFRS) in the People's Republic of China. Mortality was significantly reduced (sevenfold decrease in risk) among ribavirin-treated patients, when comparisons were adjusted for baseline risk estimators of mortality (P = .01; two-tailed). HFRS typically consists of five consecutive but frequently overlapping clinical phases. Only occurrence of oliguric phase and hemorrhage was associated with severity of clinical disease in the placebo group. Ribavirin therapy also resulted in a significant reduction in the risk of entering the oliguric phase and experiencing hemorrhage. The only ribavirin-related side effect was a well-recognized, fully reversible anemia after completion of therapy.

Double-stranded ribonuclease coinduced with interferon.

Double-stranded (ds) RNA and many viruses are inducers of interferon (IFN), the latter presumably because they contain, or can form, dsRNA. Concomitant with the induction of IFN in chicken embryo cells was the induction of a novel double-stranded ribonuclease (dsRNase), which was released into the medium and continued to accumulate long after IFN production ceased. Only avian cells (chicken, quail, turkey, or duck) expressed high levels of this dsRNase; mammalian, turtle, or fish cells did not. Production of the nuclease was inducer dose-dependent. Optimum pH and cation requirements distinguished it from other dsRNase activities. Degradation of dsRNA was endonucleolytic. Activity resided in a molecule of an Mr of approximately 34,500. Low levels of a single-stranded (ss) RNase activity were inseparable from the dsRNase. The role for a dsRNA-inducible dsRNase released from cells is unknown.

Rift Valley fever infection of rhesus monkeys: implications for rapid diagnosis of human disease.

Rhesus monkeys inoculated with Rift Valley fever (RVF) virus provide a model in which serial observations of serum viral antigen and antibodies can be made. In 9 non-fatal and 3 fatal infections, either antigen or IgM enzyme-linked immunosorbent assay (ELISA) antibodies were detected in every serum sample during the acute phase. Furthermore, viral nucleic acid could be detected by filter hybridization in most samples taken on days 1 to 3. Circulation of significant quantities of viral RNA provides an additional approach to the diagnosis and study of RVF.

Filter paper confetti in a serological Rift Valley fever survey.

In order to collect epidemiological data about the Rift Valley fever epidemic in Mauritania, we decided to use the filter paper method. The mean recovery level of specific antibodies from filter paper, tested using an immunoenzymatic method, is around one fourth. Taking the mean haematocrit into account, we estimated the extract square with a 1/300 dilution. This method was very useful for epidemiological studies, we observed few patient refusals, but it is necessary to know the exact specificity of the antibodies.

[Hemorrhagic forms of Rift Valley fever in Mauritania]. / Les formes hémorragiques de la fièvre de la Vallée du Rift en Mauritanie.

During and after a Rift Valley fever (RVF) epidemic in Southern Mauritania, we collected 600 clinical observations. 348 were confirmed to be RVF cases. Among the 5 clinical forms we observed, some are benign but others, especially those with hemorrhagic signs are serious. We observed 48 icterohemorrhagic forms with 25 deaths. An icterus was associated with hemorrhagic signs, varying from gingivorrhagia to abundant bleeding. Biological hepato-nephritis was always present in major hemorrhagic forms. Fulminant forms, spectacular and characteristic are excellent markers for epidemiological studies in the field.

[Mild clinical forms of Rift Valley fever during the epidemic in Mauritania]. / Les formes cliniques bénignes de la fièvre de la Vallée du Rift pendant l'épidémie de Mauritanie.

During and after a Rift Valley fever (RVF) epidemic in Southern Mauritania we collected 600 clinical observations. 348 were confirmed to be RVF cases. We described 5 major clinical aspects: mild, icteric, icterohemorrhagic, hemorrhagic and neurological forms. The first one is the most frequently seen with 42.8% of the cases at admission. Fever was associated with various pains (cephalalgia, myalgia, arthralgia) and an important asthenia. Inconsistently this syndrome was accompanied by epistaxis and conjunctival hyperemia. The icteric form, never described before, is an icterus occurring during evolution of a mild form. It represents 28.5% of total cases at admission. The great number of theses mild forms implies that they could be used as excellent markers for an epidemic emergence.

[Development of a clinical and biological scoring system for the prognosis of Rift Valley fever]. / Elaboration d'une grille pronostique clinique et biologique de la fièvre de la Vallée du Rift.

With regard to an acute disease, Rift Valley fever, we tried to establish a prognostic score to help physicians to set prognosis and to choose a health management suitable in their context. Using clinical and biological data collected during the 1987 RVF epidemic in Southern Mauritania, we established a prognosis score card. Data analysis allows to prognosticate forms of possible severe evolution as an association of four syndromes: fever over 39 degrees, hemorrhagic syndrome, icterus and neurological signs. Using 12 clinical symptoms and 3 biological signs, it is possible to prognosticate cases with fatal evolution.

Isolation of the Rift Valley fever virus by inoculation into Aedes pseudoscutellaris cells: comparison with other diagnostic methods.

The Rift Valley fever epidemic, which arose in the south of Mauritania beginning on October 15, 1987, enabled a comparative study of different diagnostic methods among humans. During the first two weeks of the epidemic, four parallel methods were used: inoculation into Aedes pseudoscutellaris cells, inoculation intracerebrally into suckling mice, tests by immunocapture of the circulating antigen and detection of type IgM gammaglobulins. Of 370 examined sera, 181 showed at least one marker of recent infection. Inoculation into A. pseudoscutellaris cells was by far the most sensitive and easiest method to use. Detection of the antigen by immunocapture was also a useful technique, since it allowed quick aetiological diagnosis or examination of sera conserved under poor conditions. However, its sensitivity was weak, as it could only detect 26% of positive cases. Vero cells used on a limited scale, in this particular case seemed less sensitive than A. pseudoscutellaris cells. Of a total of 991 sera, 221 diagnoses were reported by discovery of the virus and 271 by detection of specific IgM. In every case, A. pseudoscutellaris cells seemed most appropriate as the system of reference.

Assessment of an rDNA probe filter hybridization assay for the detection of Rift Valley fever virus RNA in human serum samples from the Mauritanian epidemic.

The Rift Valley fever virus (RVFV) epidemic that occurred in southern Mauritania during the 1987 rainy season provided a unique opportunity to test and evaluate a recently developed, M-segment-specific, nucleic acid filter hybridization assay on a large collection of infected human serum samples. It afforded the opportunity to compare the procedure with two other methods for detecting virus: virus isolation and antigen detection by ELISA. The filter hybridization procedure employed a polyethylene-glycol-precipitation and proteinase-K-digestion sample treatment step developed specifically for preparing serum samples for hybridization. The procedure was less sensitive for detecting RVFV in the Mauritanian human viremic samples than in sera from experimentally infected monkeys used to evaluate this procedure. It was also less sensitive than an antigen detection procedure used to test the Mauritanian samples. However, we were able to detect virus RNA in a significant proportion of the virus-isolation-positive samples. Advances in sample preparation, labelling and detection procedures, and hybridization methods will improve the sensitivity, precision and ease of use of this assay and increase its value as a diagnostic tool.

Rapid diagnosis of Rift Valley fever: a comparison of methods for the direct detection of viral antigen in human sera.

Human sera collected during the 1987 Rift Valley fever (RVF) epidemic in the Senegal River basin were analysed using three enzyme immunoassays to establish the best method for rapid diagnosis of RVF. A biotin-avidin-enhanced antigen detection method utilizing monoclonal antibodies proved most sensitive. Eighty-two viremic human sera were tested, and this assay detected antigen in 29.3% of the samples.

Rift Valley fever among domestic animals in the recent West African outbreak.

Severe haemorrhagic disease among the human population of the Senegal River Basin brought the Rift Valley fever virus (RVFV) outbreak of 1987 to the attention of science. As in previous RVFV outbreaks, local herdsmen reported a high incidence of abortion and disease in their livestock. Serum samples were obtained from domestic animal populations from areas near Rosso, the best studied focus of human infection, as well as other areas distant from known human disease. Among animals from the area of high incidence of human disease, antibody prevalence was as high as 85%, with approximately 80% of the sera positive for both RVFV IgG- and viral-specific IgM antibodies. In contrast, human populations in the same area had lower RVFV antibody prevalences, 40% or less, with 90% also being IgM-positive. Sera from livestock in coastal areas 280 km south of the epidemic area were negative for RVFV antibodies. Thus, the detection of RVFV specific IgG and IgM antibodies provided evidence of recent disease activity without the requirement to establish pre-disease antibody levels in populations or individuals and without viral isolation. Subsequently, detection of modest levels of IgG and IgM in the Ferlo region, 130 km south of the Senegal River flood plain, established that RVFV transmission also occurred in another area of the basin. Similar serological testing of domestic ungulates in The Gambia, 340 km south of Rosso, demonstrated antibody prevalence consistent with a lower level of recent transmission of RVFV, i.e., 24% IgG-positive with 6% of the positive sera also having RVFV-specific IgM.

Experimental infection of Phlebotomus papatasi with sand fly fever Sicilian virus.

Experimental studies were conducted to evaluate humans as hosts infecting the sand fly Phlebotomus papatasi with sand fly fever Sicilian (SFS) virus. Viral antigen and infectious virus circulated in the blood of infected volunteers on days 4 and 5 after intravenous inoculation with SFS virus. Viremia levels during the latter period were high enough to infect feeding sand flies, but only 13% (9/69) of the flies became infected. One out of every 3 infected sand flies that survived to feed a second time transmitted SFS to a hamster. These results confirm a vertebrate-sand fly-vertebrate transmission cycle for SFS virus, and demonstrate that horizontal transmission may contribute to the maintenance of this virus in nature.

Enzyme-linked immunosorbent assay for detection of antibodies to Rift Valley fever virus in ovine and bovine sera.

An enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of antibodies to Rift Valley fever virus (RVFV) in ovine and bovine sera. Conditions to reduce nonspecific reactions were optimized. The ELISA results correlated with those of a plaque-reduction neutralization test, revealing 100% specificity and 90.7% sensitivity. In sera from sheep and cattle inoculated against RVFV, the hemagglutination-inhibition test in combination with the ELISA provided a better indication of response to killed RVFV vaccine than did either test alone.

Comparison of an indirect fluorescent-antibody test with an enzyme-linked immunosorbent assay for serological studies of Lyme disease.

An enzyme-linked immunosorbent assay was compared with an indirect fluorescent antibody test for its ability to detect antibodies to the Lyme disease spirochete in sera of naturally infected humans, dogs, and white-footed mice and experimentally infected Swiss mice. Ninety-five percent of the total 123 sera analyzed reacted similarly in both tests. For 36 human sera, the correlation coefficient (r = 0.47) for logarithmic transformations of indirect fluorescent antibody and enzyme-linked immunosorbent assay titers was significant at P less than 0.01. Within each mammalian species, mean titers for indirect fluorescent antibody and enzyme-linked immunosorbent assay antibodies were within three-fold. Comparisons of different naturally infected mammals revealed relatively higher average titration endpoints in both tests for patients with Lyme disease. Human sera also had the widest range of titers. Both methods proved satisfactory for serological confirmation of prior spirochetal infections.

Use of enzyme immunoassays and the latex agglutination test to measure the temporal appearance of immunoglobulin G and M antibodies after natural infection or immunization with rubella virus.

The time course of appearance of antibodies after infection with rubella virus was determined with an immunoglobulin G (IgG) detection enzyme-linked immunosorbent assay, a latex agglutination test, and an IgM detection enzyme-linked immunosorbent assay. In six naturally infected rubella patients and 26 vaccinees, antibodies measured by either the IgG enzyme-linked immunosorbent assay or the latex agglutination test generally appeared in parallel with those detected by the hemagglutination inhibition test. By 28 days after inoculation of live virus vaccine and by 2 days postonset of clinical rubella symptoms caused by natural infection, antibodies were found by the two tests for all individuals. A commercially available enzyme-linked immunosorbent assay kit was used to detect rubella-specific IgM. After natural infection, IgM appeared earlier than IgG, and although IgM titers decreased rapidly postinfection, in four of five patients antibodies were still detectable 40 to 43 days after the onset of clinical symptoms. After vaccine-induced infection, rubella-specific IgM was lower in titer than after natural infection and was detected in only three of seven vaccinees 70 days post-immunization.

Comparison of the latex agglutination test with the hemagglutination inhibition test, enzyme-linked immunosorbent assay, and neutralization test for detection of antibodies to rubella virus.

The ability of a rapid, latex agglutination test to diagnose rubella infection and to measure immune status was evaluated by comparison with the hemagglutination-inhibition (HAI) test, enzyme-linked immunosorbent assay (ELISA), and the neutralization (NT) test. The latex agglutination test accurately detected serological conversions in 74 pairs of sera representing 21 natural infections and 53 immunizations. The antibody levels of 276 sera from the general population were determined by latex agglutination, HAI, and ELISA. The correlation coefficients between the titers obtained by HAI and latex agglutination and by ELISA and latex agglutination were statistically significant. Results on 12 sera did not agree when measured by the three tests. These sera were included among the 196 specimens tested by NT. The correlation coefficient between NT and latex agglutination titers was statistically significant. There was one serum positive by latex agglutination but negative by NT, and five sera were negative by latex agglutination but had titers of 4 to 8 in the NT. The relative sensitivity of detecting antibody was greater by latex agglutination than by HAI. An additional 49 sera containing residual nonspecific hemagglutinin inhibitors were evaluated by latex agglutination and NT. The untreated sera showed no false positive reactions, and 36 of 39 NT positive sera were positive in the latex agglutination test.

Detection of La Crosse arbovirus antigen in mosquito pools: application of chromogenic and fluorogenic enzyme immunoassay systems.

An enzyme immunoassay producing either a chromogenic or fluorogenic end product was developed and evaluated for detecting La Crosse viral antigen within mosquito pools. The enzyme immunoassay was found to be sensitive, detecting one infected mosquito within a pool of 100 mosquitoes, and specific, distinguishing between closely related California group viruses. Assays were completed within 5 h after the addition of test samples. La Crosse viral antigen could be readily detected in mosquito pools after seven freeze-thaw cycles.