Resistance of mice with experimental histoplasmosis to infection with Rickettsia typhi.
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A seroepidemiology study was done in response to the recent increase of Endemic Typhus cases diagnosed at University Hospital. The serosurvey was based on doctors' request for the Weil Felix (WF) or the Indirect Immunoperoxidase (IIP) test in Pyrexia of Unknown Origin (PUO) patients for the years 1991 to 1997. Over the 7 years, we found that the incidence of Endemic typhus is increasing with gender (male:female = 2:1), age (20-40 years) and race distribution (Indians > Malay > Chinese) that reflects socioeconomic circumstances. A commercially available ELISA dot assay [INDX (E2R3) Dip-S-Ticks], for the detection of antibodies against R. typhi was compared with the indirect immunoperoxidase test (IIP). The ELISA assay was done against 219 IIP tested sera. The Dip-S-Ticks was found to be comparable to the IIP with a sensitivity of 91.7% and specificity of 92.8% at cut-off titres of > 1:80 IIP.
Rickettsial diseases except those belonging to spotted fever group rickettsioses are poorly studied in South America particularly in Brazil where few epidemiological reports have been published. We describe a serosurvey for Rickettsia rickettsii, R. typhi, Coxiella burnetii, Bartonella henselae, B. quintana, and Ehrlichia chaffeensis in 437 healthy people from a Brazilian rural community. The serum samples were tested by indirected micro-immunoflourescence technique and a cutoff titer of 1:64 was used. The seroprevalence rates for R. rickettsii, R. typhi, C. burnetii, B. henselae, B. quintana, and E. chaffeensis were respectively 1.6% (7 samples); 1.1% (5 samples); 3.9% (17 samples); 13.7% (60 samples); 12.8% (56 samples), and 10.5% (46 samples). Frequent multiple/cross-reactivity was observed in this study. Age over 40 years old, urban profession, and rural residence were significantly associated with some but not all infections rate. Low seropositivity rates for R. rickettsii, R. typhi, and C. burnetii contrasted with higher rates of seropositivity for B. quintana, B. henselae, and E. chaffeensis. These results show that all tested rickettsial species or antigenically closely related possible exist in this particular region.
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Paired sera from cases of epidemic typhus in Ethiopia and from probable cases of Rocky Mountain spotted fever (RMSF) in the United States were examined by microagglutination (MA) and microimmunofluorescence (micro-IF) tests for antibodies against Rickettsia prowazekii, Rickettsia typhi, Rickettsia canada, Rickettsia rickettsii, Rickettsia conorii and Ricksettsia akari. IgG and IgM antibodies against the various rickettsiae were titrated with specific fluorescein-conjugated anti-IgG and anti-IgM sera. Purified, particulate rickettsial antigens were employed in all tests. A majority of patients acutely ill with epidemic typhus produced both IgG and IgM antibodies against R. prowazekii, R. typhi and R. canada. Concurrently they produced IgG (but seldom IgM) antibodies against members of the spotted fever group. In contrast, patients ill with probably spotted fever, while producing IgG and IgM antibodies against R. rickettsii, R. conorii and R. akari, also produced both IgG and IgM antibodies with about equal frequency against members of the typhus group. It was concluded that a relatively broad antigenic relationship exists between rickettsiae of the typhus and spotted fever groups.
A microplate enzyme-linked immunosorbent assay (ELISA), developed for the detection of antibodies to typhus group rickettsiae, was used to analyze human sera from individuals engaged directly or indirectly in rickettsial research. The earliest serum available from each of 112 individuals was tested for immunoglobulin M (IgM) and IgG antibodies against Rickettsia typhi and Rickettsia prowazekii by ELISA at a 1:500 dilution. In at least one assay, nine sera had ELISA optical densities of greater than 0.2, which were above the mean optical densities plus three standard deviations of the other 103 sera. Three of the positive sera were from individuals with known clinical cases of typhus infection. The other sera with predominantly IgG titers were from individuals with extended laboratory exposure to rickettsiae or histories of typhus vaccination, or both. During continued serological surveillance, eight additional people with repeated occupational exposure to typhus rickettsiae had seroconversions in the ELISA to optical densities of greater than 0.2. No apparent clinical illness occurred in two individuals, whereas six clinical cases of infection occurred in others subsequent to accidental laboratory autoinoculation (one) or aerosol exposures (five). In the clinical infections, antibodies were first detected at 7 days, but in subsequent sera, rises and declines in titers were quite variable and were influenced by vaccination, relapse, and time and extent of antibiotic therapy. In primary infections the sera of several individuals who received immediate antibiotic therapy had brief strong IgM responses without pronounced increases in IgG. In contrast, much higher IgG levels were attained in three cases in which relapse occurred, the individual had previously been immunized, or treatment had been delayed. The microplate ELISA proved to be a highly sensitive and reliable test for detection of the human serological response to typhus antigens.
An indirect hemagglutination (IHA) test is described that uses glutaraldehyde-stabilized erythrocytes treated with a rickettsial erythrocyte-sensitizing substance obtained from Rickettsia typhi or Rickettsia rickettsii. The serological reagent was stable for at least 3 months at room temperature and 6 months at 4 C. It exhibited group specificity and no group cross-reactivity. At a minimum dilution of 1:40, acute and early convalescent epidemic and murine typhus antisera showed 86% positive reactors, whereas similar spotted fever antisera had 74% positive reactors. In comparison with the indirect fluorescent antibody test, the IHA procedure gave lower titers but showed comparable detection of seroconversion with most paired sera. The IHA test demonstrated significantly higher titers than the complement fixation test and was more sensitive than either the complement fixation or Weil-Felix test in identifying seroconversion. No agglutination was observed when sensitized erythrocytes were tested with rodent sera known to contain rickettsial antibodies.
Primer pairs for PCR were designed from the gene encoding the 17,000-molecular-weight genus-common antigen of Rickettsia japonica, Rickettsia rickettsii, Rickettsia conorii, Rickettsia typhi and Rickettsia prowazekii. Primers R1, R2 were designed for amplifying the genomic DNA from spotted fever group (SFG) rickettsiae and epidemic typhus rickettsiae. Primers Rj5, Rj10 were designed for amplifying the genomic DNA from only R. japonica. Using the primers R1, R2, about a 540-bp fragment was observed by amplifying the genomic DNA from R. japonica, R. rickettsii, R. conorii, Thai tick typhus TT-118, Rickettsia sibirica, Rickettsia montana, Rickettsia askari, R. typhi, R. prowazekii and Katayama strain isolated from the patient infected with SFG rickettsiae. Using the primers Rj5, Rj10, the 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and Katayama strain. Therefore, the Katayama strain was identified to belong to R. japonica. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp bands were amplified from blood of the patients infected with SFG rickettsiae in Kanagawa prefecture. These findings indicate that the causative agent of SFG rickettsiosis in these two patients was R. japonica. The ticks, Ixodes ovatus and Haemaphysalis flava, were collected by out field research in Kanagawa prefecture. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp were amplified from these ticks. This indicates that I. ovatus and H. flava were the vector of R. japonica in Kanagawa prefecture. Also, with the primers R1, R2, about a 540 bp fragment was amplified but with primers Rj5, Rj10, no fragments were amplified from I. ovatus and H. flava. Therefore, these ticks may have SFG rickettsiae other than R. japonica and epidemic typhus rickettsiae.
A study was conducted between 1984 and 1987 to determine the prevalence of Rickettsia typhi and Rickettsia conorii infections among humans residing in the Nile Delta, Suez Canal area and Nile Valley of Egypt. Serum specimens were obtained from garbage and rodent control workers, other unclassified occupational workers, and from patients with fever of undetermined aetiology. All sera were assayed for IgA + IgM + IgG (IgAMG) antibody mixture and if positive, reassayed for specific IgM antibody to rickettsia by the indirect fluorescent antibody technique. R. typhi antibody was found in 19% (33/178) of the garbage collectors, whereas only 1% (2/178) had demonstrable antibody to R. conorii. Among those with other occupations, R. typhi antibody was detected in 0.7% (2/295) and none had R. conorii antibody. The antibody prevalence rate for R. typhi among patients with febrile illness ranged from 25 to 41%, and from 2 to 15% for R. conorii, at three different locations in Egypt. In addition, IgM antibody to R. typhi was demonstrated in some patients showing symptoms compatible with rickettsial disease and in some patients who seroconverted, indicating that R. typhi was the cause of illness among some of these patients. These findings support previous observations that R. typhi and R. conorii are the causes of human rickettsial disease in Egypt, and that humans are commonly infected with R. typhi.