Hantavirus Serotype Research Articles

Development of monoclonal antibodies against Hantaan virus nucleocapsid protein.

Forty-five hybridoma cell lines producing monoclonal antibodies against Hantaan virus, the etiologic agent of hemorrhagic fever with renal syndrome, were generated by fusion of P3-X63-Ag8.V653 myeloma cells with spleen cells of mice immunized with inactivated Hantaan virus vaccine. Among these, 38 antibodies were identified as binding to the 48-kDa nucleocapsid protein by immunoblot assay or radioimmunoprecipitation. Twenty-six of them were of the immunoglobulin G1 (IgG1), nine were of the IgG2a, and three were of the IgA isotype. According to cross-reactivities with other serotypes of the genus Hantavirus, the antibodies were classified into three groups: 6 antibodies specific to the Hantaan serotype (group I), 20 antibodies cross-reacting with Hantaan and Seoul serotypes (SR-11, Tchoupitoulas, and R22) (group II), and 12 antibodies cross-reacting with Hantaan, Seoul, and Prospect Hill serotypes (group III). None of the antibodies cross-reacted with the Puumala serotype. With a panel of antibodies of different cross-reactivities, serotypes of Hantavirus could be differentiated. Thirty-eight monoclonal antibodies against Hantaan virus nucleocapsid protein which have different cross-reactivities between serotypes were developed. These results confirmed the presence of multiple serotype-specific epitopes on the nucleocapsid protein of Hantaan virus, which can be utilized in differentiation of serotypes.

Hantavirus Pulmonary Syndrome: The Four Corners Disease

To review the epidemiology, pathogenesis, clinical features, diagnosis, and treatment of hantavirus infections, focusing on the recent outbreak of hantavirus pulmonary syndrome in the US. A MEDLINE search (1966 to present) of English language literature pertaining to hantaviruses was performed. Additional literature was obtained from reference lists of pertinent articles identified through the search. All articles were considered for possible inclusion in the review. Pertinent information, as judged by the authors, was selected for discussion. Hemorrhagic fever with renal syndrome (HFRS) has long been recognized in Eurasia and is the predominant disease manifestation of hantavirus infection worldwide. Hantavirus pulmonary syndrome (HPS) recently has been described in the US and exhibits greater pulmonary involvement and mortality than HFRS. Historically, 4 hantavirus serotypes (Hantaan, Seoul, Puumala, Prospect Hill) are recognized; however, additional serotypes have been proposed as distinct serogroups, including the serotype responsible for HPS in the Four Corners area: the Four Corners virus (FCV). Phylogenetic analysis shows that FCV is most closely related to Prospect Hill virus, another hantavirus previously isolated in the US that has not yet been identified with human disease. Additional hantavirus serotypes isolated in the US may provide insight into the prevalence of hantavirus infection and disease in this country. Inhalation of aerosolized virus is the predominant mechanism of hantavirus infection. Diagnosis is based primarily on clinical findings and serologic evidence of hantavirus antibody or direct evidence in clinical tissue specimens. Limited clinical studies evaluating ribavirin as a therapeutic modality demonstrated that the agent improves clinical outcome in HFRS. However, the role of ribavirin in the treatment of HPS remains to be determined. Hantavirus infections are becoming increasingly recognized as a cause of disease worldwide. Recognition of hantavirus disease in the US suggests enzoonosis of pathogenic hantaviruses. In the absence of a well-established cure, early diagnosis is imperative so that aggressive supportive care can be initiated.

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. It poses a special danger to humans who have close contact with field rodents.

Application of a Recombinant Baculovirus Expressing Hantavirus Nucleocapsid Protein as a Diagnostic Antigen in IFA Test: Cross Reactivities among 3 Serotypes of Hantavirus which Causes Hemorrhagic Fever with Renal Syndrome(HFRS).

Recombinant baculovirus-infected insect cells which expressed recombinant protein, analogous to the nucleocapsid protein (NP) of Seoul type hantavirus, strain SR-11 (rNP-SR-Sf9) were applied to the indirect immunofluorescent antibody (IFA) test. The rNP-SR-Sf9 reacted with anti NP MAb clones which recognized strain specific or hantavirus common epitopes in the IFA test. The recombinant antigen was detected by antibodies to 3 serotypes of hantaviruses (Hantaan 76-118, SR-11, Puumala). Antibody titers of a group of experimentally infected mouse and urban rat sera using rNP-SR-Sf9 and strain SR-11-infected Vero cell antigens were mutual correlative. These results indicated that the rNP-SR-Sf9 were an effective and safety substitute for Vero E6 cells in the IFA test for serosurveys of hantavirus infection.

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. The RNA of this virus was successfully amplified with hantavirus genus reactive primer sets by reverse transcriptase polymerase chain reaction (RT-PCR); however, PCR-RFLP analysis of the amplified product was shown to be unique among those of the known hantaviruses, further indicating that Dobrava virus represents a new hantavirus serotype.

Distribution of hantavirus serotypes Hantaan and Seoul causing hemorrhagic fever with renal syndrome and identification by hemagglutination inhibition assay

An epidemiologic evaluation of patients with hemorrhagic fever with renal syndrome from different locations in the People's Republic of China was conducted to define the prevalence of two Hantavirus serotypes, Seoul (SEO) and Hantaan (HTN). Serum specimens were collected between 5 and 14 days after the onset of illness and were tested for antibodies by both hemagglutination inhibition (HI) and plaque reduction neutralization (PRN). By the HI test, the geometric mean titer (GMT) of antibodies to SEO in the sera from individuals from Kaifeng City of Henan Province was five times higher than that to HTN. In contrast, by the HI test, the sera from individuals from Jiande County of Zhejiang Province had a GMT of antibodies to HTN that was seven times higher than that to SEO. In the sera from individuals from Shanghai, only a twofold difference was observed in HI antibody titers to the two hemagglutinins by the HI test, with that to HTN being higher than that to SEO. By the PRN test, the GMT ratios of antibody between HTN and SEO strains from individuals in Kaifeng, Jiande, and Shanghai were found to be 1:13, 14:1, and 2:1 respectively. A close correlation (r = 0.8219) and concordance rate (78.3%) were observed between the PRN and HI tests for the identification of the serotypes of individual cases of hemorrhagic fever with renal syndrome. The hantavirus serotypes from individuals in Kaifeng and Jiande were identified as predominantly SEO and HTN, respectively, and those from individuals in Shanghai had an indeterminant serotype defined by these two techniques. The HI test appears to be a simple and reliable way of determining the predominant hantavirus that causes HFRS in a given geographic area.

Identification of hantavirus serotypes by testing of post‐infection sera in immunofluorescence and enzyme‐linked immunosorbent assays

Serum samples were collected from 27 individuals who had been infected with a member of the genus Hantavirus in the Netherlands or Belgium during the last 15 years. These samples were tested in an immunofluorescence assay (IFA) and two enzyme-linked immunosorbent assay (ELISA) systems, using different virus strains that represented each of the four recently proposed serotypes of this genus. The serum samples from 11 individuals who had been infected through contacts with laboratory rats showed the highest reactivities with Hantaan virus (serotype I) and SR-11 (serotype II) in the IFA and ELISA systems. The samples of 16 individuals who had probably been infected through contacts with wild rodents showed the highest reactivities with Hällnäs virus (serotype III) in the IFA. All except two of these also showed the highest reactivity with Hällnäs virus in the two different ELISA systems.

Hemorrhagic Fever with Renal Syndrome in Yugoslavia: Antigenic Characterization of Hantaviruses Isolated from Apodemus Flavicollis and Clethrionomys Glareolus

Hantavirus antigens were detected in lung tissues of 8/113 Apodemus flavicollis and 2/17 Clethrionomys glareolus captured in 1984 in Fojnica, a region of Yugoslavia endemic for hemorrhagic fever with renal syndrome; hantavirus antigens were not detected in lung tissues from 126 other mammals collected in Fojnica. Three hantaviruses, 2 from A. flavicollis and 1 from C. glareolus, were isolated directly in Vero E6 cells and were partially characterized. The isolates from A. flavicollis, designated Fojnica virus, were antigenically similar but not identical to Hantaan virus strain 76-118, whereas the isolate from C. glareolus was antigenically indistinguishable from Puumala virus, strain Hällnäs B1. These data are consistent with previous studies that indicate the existence of at least 2 hantavirus serotypes in Yugoslavia.

Hemorrhagic fever with renal syndrome in Yugoslavia: detection of hantaviral antigen and antibody in wild rodents and serological diagnosis of human disease.

Lung tissues from 547 rodents and 26 insectivores captured between 1981 and 1984 in central Bosnia (Fojnica) and central Serbia (Cacak), 2 regions known to be endemic for hemorrhagic fever with renal syndrome (HFRS), were examined for hantaviral antigen by the indirect immunofluorescent antibody technique. Antigen was detected in 17/231 Apodemus flavicollis, 3/187 A. sylvaticus, 1/46 A. agrarius, 4/32 Mus musculus, and 3/28 Clethrionomys glareolus. In addition, antibodies against Hantaan and Puumala viruses were found in serum pooled from 2 C. glareolus captured in Fojnica and 6 Pitimys subterraneus caught in Cacak. Sera of 27 HFRS patients from different parts of Yugoslavia were tested against 3 serotypes of hantavirus. Patients from Bosnia and Serbia had highest titers against Hantaan virus, while patients from Croatia had highest titers against Puumula virus, the agent of nephropathia epidemica.