Serological diagnosis of hantavirus infections by an enzyme-linked immunosorbent assay based on detection of immunoglobulin G and M responses to recombinant nucleocapsid proteins of five viral serotypes.

Worldwide, hantaviruses cause more than 100,000 human infections annually. Rapid and accurate methods are important both in monitoring acute infections and for epidemiological studies. We and others have shown that the amino termini of hantavirus nucleocapsid proteins (Ns) are sensitive tools for the detection of specific antibodies in hantavirus disease.

The hantaviruses of Europe: from the bedside to the bench.

In Europe, hantavirus disease can hardly be called an emerging zoonosis; it is rather a rediscovered disease. Since 1934 an epidemic condition with primarily renal involvement has been described in Sweden. Nowadays, hundreds to thousands of cases per year are registered in Fennoscandia, fluctuating with the numbers of the specific Arvicoline-rodent reservoir, the red bank vole, which carries the main European serotype, Puumala (PUU).

Tick-borne encephalitis: possibly a fatal disease in its acute stage. PCR amplification of TBE RNA from postmortem brain tissue.

Tick-borne encephalitis has occurred regularly in Europe since it was first diagnosed in 1931 by Schneider. The mortality rate of patients with this disease is 1-2%. Death usually occurs in the acute stage of illness.

Genetic and antigenic properties of Dobrava virus: a unique member of the Hantavirus genus, family Bunyaviridae.

We examined the genetic and antigenic properties of Dobrava (DOB) virus, a hantavirus associated with severe haemorrhagic fever with renal syndrome in Europe. Cloning and sequence analyses revealed the DOB M segment to consist of 3644 nucleotides, with a coding capacity of 1134 amino acids in the virus complementary-sense RNA (cRNA). Seven potential asparagine-linked glycosylation sites were identified in the M segment gene product, one in the G2 and six in the G1 coding regions.

Laboratory acquired tick-borne meningoencephalitis: characterisation of virus strains.

Background: The handling of tick-borne encephalitis (TBE) virus is potentially hazardous, as indicated by a number of laboratory-acquired infections in the prevaccination era.

Objectives: (1) To reemphasize the hazard of handling TBE virus without being vaccinated by describing the case of a laboratory-acquired full-blown TBE in a microbiologist who isolated the virus from a blood sample. (2) To molecularly characterize the causative virus strain isolated in Slovenia in comparison with the European prototype strain Neudoerfl.

Evaluation of two commercial amplification assays for detection of Mycobacterium tuberculosis complex in respiratory specimens.

To evaluate the usefulness of two standardized commercially available amplification assays for the detection of Mycobacterium tuberculosis: Amplicor test (Roche) and MTD-Amplified direct test (Gen-Probe) a total of 281 respiratory specimens from 198 patients with symptoms of pulmonary diseases were examined and compared with conventional methods. Fifty-seven specimens were positive and 218 negative by both amplification assays. Three specimens were reactive by Amplicor only, and three by MTD only.

Isolation of DNA from archival Papanicolaou stained cytological smears using a simple salting-out procedure.

DNA from archival Papanicolaou stained and unstained cytological smears was successfully isolated using a simple, rapid and inexpensive salting-out procedure. The quality of DNA was controlled by polymerase chain reaction (PCR) amplification of segments of the human beta-globin, human beta-actin and human papillomavirus L1 genes. Only negligible differences in amplification efficiency were observed between DNA isolated from stained and unstained smears.

Isolation of a strain of a Hantaan virus from a fatal case of hemorrhagic fever with renal syndrome in Slovenia.

Hemorrhagic fever with renal syndrome (HFRS) has been serologically confirmed in Slovenia during the last seven years. There is evidence that three hantaviruses (Hantaan, Puumala, and a newly described form termed Dobrava) circulate simultaneously in this area. Recently, a hantavirus was isolated from the urine and brain tissue of a fatal case of HFRS.

Belgrade virus, a cause of hemorrhagic fever with renal syndrome in the Balkans, is closely related to Dobrava virus of field mice.

Belgrade virus is a recently described hantavirus that causes severe hemorrhagic fever with renal syndrome (HFRS) in people living in various parts of the Balkan Peninsula. Nucleotide sequencing of the G2-encoding region in the medium (M) segment of the viral genome, reverse transcribed and amplified by the polymerase chain reaction, revealed the Belgrade virus to be substantially different from Hantaan virus and other major serotypes of hantavirus but identical to Dobrava virus, a virus isolated from a field mouse (Apodemus flavicollis) in Slovenia. Belgrade virus may be an important cause of HFRS in the Balkan Peninsula, extending north toward the Alps.

Dobrava virus as a new Hantavirus: evidenced by comparative sequence analysis.

Dobrava virus, recently isolated from a yellow-neck mouse (Apodemus flavicollis), captured in a northern Slovenian village where severe cases of hemorrhagic fever with renal syndrome were recognized, was shown by serology and restriction enzyme digestion of PCR-amplified gene segments to be related to previously recognized hantaviruses. To investigate further the relationship of this new isolate to other hantaviruses, a portion of the medium (M) genome segment of Dobrava virus was amplified by PCR and the nucleotide sequence determined. Comparing the nucleotide sequence with the same gene region of other hantaviruses revealed an overall homology of 41.

Characterization of Dobrava virus: a Hantavirus from Slovenia, Yugoslavia.

Small mammals were collected in natural foci of hemorrhagic fever with renal syndrome (HFRS) in Slovenia, Yugoslavia, and a hantavirus was isolated from the lungs of an Apodemus flavicol lis captured in Dobrava village. This new isolate, Dobrava virus, was compared with representative strains of the Hantavirus genus by serological and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. It was found by cross immunofluorescent and enzyme-linked immunosorbent assays that antigenic properties of Dobrava virus were different from those of other hantaviruses.

Evidence for Hantavirus disease in Slovenia, Yugoslavia.

In Slovenia, North-Western part of Yugoslavia, 17 clinically documented Hantavirus disease cases (HVD) were serologically confirmed so far. Previously HVD was reported in the Southern part of Yugoslavia. By the indirect fluorescent antibody test (IFA), the prevalence of IgG class antibodies against different Hantaviral antigens was demonstrated in human sera collected in Slovenia.