Detection of anti-Coxiella burnetii antibodies by immunofluorescent technique and enzyme-linked immunosorbent assay.
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We evaluated real-time PCR assays for the detection of C. burnetii which targets sequences that are present either in one (icd) or in several copies (transposase of IS1111a) on the chromosome. The assays are highly sensitive, with reproducible detection limits of approximately 10 copies per reaction, at least 100 times more sensitive than capture ELISA, when performed on infected placenta material and specific for C. burnetii. The numbers of IS1111 elements in the genomes of 75 C. burnetii isolates were quantified by real-time PCR and proved to be highly variable.
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The dramatic spread of Q fever in Poland among cattle kept in isolation from natural environment (ticks, wild animals) has suggested the possibility that the infection may also be transmitted sexually. To test this hypothesis series of experiments have been performed in controlled laboratory conditions. Male mice infected with C. burnetii were allowed to mate with healthy female mice. On day 18 of pregnancy serum IgM antibodies to C. burnetii antigens and bacteria in spleen, liver and placenta were detected. The influence of C. burnetii transmission between parents of their offspring was investigated. It has been found that C. burnetii infection in males diminish the number of fertilized females. Their litters are fewer in number and the number of dead embryos is increased.
This study investigates the complete genome sequences of the Nine Mile, K, and G isolates to improve our understanding of the unique virulence properties of C. burnetii and to provide important insight into the genome architecture and genetic diversity of this pathogen.
Serological examination of bovine and human sera for antibodies against Coxiella burnetti was carried out by the immunofluorescence technique. Twenty to 30% of the cows examined were antibody-positive. Sera from two veterinarians also had antibody against C. burnetii. These results suggest an increase in the number of infected cows with C. burnetii in Japan since 1954, and also imply the possibility of the prevalence of acute Q fever in the human population, which had been underestimated and undiagnosed for the last three decades.
The authors developed monoclonal antibodies to Coxiella burnetti, the agent of Q fever. The selected monoclonal antibody, Cox1D8, did not cross-react with other bacteria and was used for early detection of C burnetti in shell vial cell cultures and for staining C burnetii in paraffin embedded tissues. Formalin or Bouin fixation did not alter the reactivity of the antigen with the antibody. This monoclonal antibody could be useful in the pathologic diagnosis of Q fever hepatitis and endocarditis.
Biological properties of antigens of whole cells of C. burnetii in phase I (WCI), their residue (CMR) and extract (CME) obtained by extraction of C. burnetti with chloroform/methanol mixture were subjected to a comparative study. It was shown that CME is able to react specifically with sera containing antibodies against WCI. The inoculation of animals with CME, but not with WCI and CMR does not cause changes of humoral and cell immunity to Coxiella and does not strengthen nonspecific antibacterial activity, but it can stimulate the formation of antibodies. Morphological changes of internal organs of mice were most expressed after the CMR inoculation and were practically absent after the CME inoculation.
The DNA from the Nine Mile phase I strain of Coxiella burnetti, the etiological agent of Q fever, has been isolated and purified by cesium chloride-ethidium bromide density gradient centrifugation. A fraction of this DNA has a density characteristic of plasmid DNA. The plasmid DNA was cut with 20 different restriction endonucleases and shown to be a discrete entity. The plasmid, designated QpH1, is approximately 36 kilobases in size and has a molecular mass of 2.4 x 10(7) daltons. A partial restriction map of QpH1 has been constructed by using the restriction endonucleases SalI, KpnI, PstI, and XbaI. QpH1 DNA radioactively labeled by nick translation was used to show that sequences similar to the plasmid are also present in the phase II antigenic variant of C. burnetii.
In this paper, we describe the identification of an iron-acquisition gene homologue, ferrous iron transporter (feoB) in C. burnetii. The results of a genomic screen for putative Fur-regulated genes and Fur boxes and the development of a two-plasmid system to analyze these Fur boxes will also be illustrated.
As found out in earlier investigations, after the intratracheal infection of guinea pigs with C. burnetii (the method of infection, most similar to aerogenic infection which is characteristic of Coxiella under natural conditions) the maximum amount of the infective agent is accumulated in the lungs of the animals, damaging the organs of the reticuloendothelial system. The present investigation has revealed that this method of infection makes it possible differentiate C. burnetii virulent strains, differing in the level of the accumulation of the infective agent in the lungs of guinea pigs.
Four commercially available serological assays for the detection of IgM phase II antibodies in patients with acute Q fever infection were compared using a panel of 23 serum samples from patients with acute Q fever and 88 control sera from blood donors.
Intracellular pathogens customize the composition and function of the vacuoles they occupy, and can arrest or distort vacuolar maturation. In doubly infected cells, vacuoles that contain two different parasites can be used to test for exclusionary mechanisms, for expression of vacuolar phenotypes that permit or restrict fusion, and for the survival of pathogens targeted to an unusual cellular compartment.
Q Fever has proved to be a virulent infectious disease for a great many years. A number of epidemics have occurred among laboratory workers engaged in research on C burnetii but few fatalities have been recorded and the course of the disease is generally uncomplicated and benign (Braunwald et al, 1987). By following the recommendations of a large number of scientists, implementing and maintaining basic control measures, the risk of contamination will be kept at a minimum.
Granulomatous hepatitis as the sole manifestation of acute Q fever has been reported only rarely, although minimal hepatic dysfunction may be common in the acute disease. In this paper we report two patients with acute Q fever who presented with hepatitis; one of whom had granulomatous hepatitis on liver biopsy. We discuss the serological diagnosis of acute and chronic Q fever particularly in relation to hepatitis.