OBJECTIVESTo develop and evaluate a method for detection of the total antibodies against hemorrhagic fever with renal syndrome (HFRS) virus with improved sensitivity and simplified operation procedure.

METHODSThe nucleic proteins of hantavirus were used as coating antigens as well as detection antigens labeled with horse radish peroxidase (HRP). The operation protocol was established, optimized and compared with indirect fluorescence assay (IFA).

RESULTSThe specificity of this method was 100 percent in the test of different human sera and 4-8 times more sensitive than IFA. And, it is simpler without requiring any change of the reagents, different sources of samples did not affect the results of the test.

CONCLUSIONThis method is specific, sensitive and simple for detection of the total antibodies in sera against hantavirus, could be used for the screening of Hantavirus infection in human and host rodent animals.

Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; methods ; Hantaan virus ; immunology ; Humans ; Reproducibility of Results ; Sensitivity and Specificity

Hantaan virus ; genetics ; isolation & purification ; Hemorrhagic Fever with Renal Syndrome ; prevention & control ; virology ; Humans ; Viral Vaccines ; immunology

The efficacy of a formalin-inactivated hemorrhagic fever with renal syndrome (HFRS) vaccine and the effectiveness of a related vaccination program have not been previously evaluated. We measured the primary immune responses to Hantavax by plaque reduction neutralizing antibody test (PRNT), hemagglutination inhibition test (HAI), ELISA and high density particle agglutination test (HDPA) in order to confirm a possible biological efficacy through independent substantiation of experimental results and to compare the results with previous studies. Following two doses of primary vaccination, the seroconversion rate of PRNT and HAI antibody was 33.3% (10/30)[95% C.I. 17.3-52.5%] and 26.7% (8/30) [95% C.I. 12.3-45.9%], respectively. The correlation between PRNT and HAI antibody showed a statistical significance (r=0.58, p 0.01). The seroconversion rate of HDPA and ELISA were both 76.7% (23/30) [95% C.I. 57.7-90.1%], which correlated well with each other(r=0.58, p 0.01). In our study, Hantavax elicited low neutralizing antibody responses, at least in the volunteers samples that we tested. The vaccination program, including the vaccine itself, that has been adopted by the national immunization program to protect against HFRS in Korea should be re-evaluated and re-formulated to produce a higher protective immune response rate.
Adult ; Antibodies, Viral/*blood ; Hantaan Virus/*immunology ; Human ; IgG/blood ; Korea ; Middle Age ; Time Factors ; Vaccination ; Vaccines, Attenuated/immunology ; Viral Vaccines/*immunology

Animals ; China ; epidemiology ; Disease Reservoirs ; Hantaan virus ; immunology ; isolation & purification ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; prevention & control ; transmission ; Humans ; Prevalence ; Vaccination ; Vaccines, Inactivated ; Viral Vaccines ; immunology

OBJECTIVETo adapt the candidate strains of hemorrhagic fever with renal syndrome (HFRS) purified vaccine to Vero cells and to study their antigenicity and immunogenicity.

METHODSThe viral strains H8207 (Hantaan virus, HTN) and Y86013 (Seoul virus, SEO) were continuously propagated in Vero cell by the terminal dilution method and studied the characteristics of virus multiplication, viral titers and the amounts of virus antigen after serial passages. Three batches of crude monovalent inactivated vaccine were developed using the different passages of these 2 viral strains.

RESULTSThe strains H8207 and Y86013 adapted to Vero cells and stably grew on the cells with high titers. Rabbits immunized with the crude vaccines of H8207 and Y86013 showed 100% sero-conversion and the neutralizing antibody titers of the rabbit immune sera reached 1?10 at 4 weeks after 2 times of immunization.

CONCLUSIONSThe results suggest that these 2 candidate strains had adapted to Vero cells, possessed high titers and good immunogenicity and be feasible to prepare the HFRS purified vaccine in Vero cells.

Animals ; Antibodies, Viral ; blood ; Cercopithecus aethiops ; Hantaan virus ; growth & development ; immunology ; Hemorrhagic Fever with Renal Syndrome ; prevention & control ; Mice ; Neutralization Tests ; Rabbits ; Seoul virus ; growth & development ; immunology ; Vaccination ; Vaccines, Inactivated ; immunology ; Vero Cells ; Viral Vaccines ; biosynthesis ; immunology

This is the first case of virus-associated encephalitis/encephalopathy in which the pathogen was Hantaan virus. A 53-yr-old man presented fever, renal failure and a hemorrhagic tendency and he was diagnosed with hemorrhagic fever with renal failure syndrome (HFRS). In the course of his illness, mild neurologic symptoms such as dizziness and confusion developed and magnetic resonance images revealed a reversible lesion in the splenium of the corpus callosum. This case suggests that HFRS patients with neurologic symptoms like dizziness and mental slowing should be considered to have structural brain lesions and to require brain imaging studies.
Antibodies, Viral/blood ; Corpus Callosum/*pathology ; Diagnosis, Differential ; Hantaan virus/immunology ; Hemorrhagic Fever with Renal Syndrome/*diagnosis/therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Platelet Count ; Renal Dialysis

OBJECTIVETo evaluate the effect the vaccine of hemorrhagic fever with renal syndrome (HFRS).

METHODSData of totally, 1095 individuals who were immunized with HFRS vaccine type produced by Shenyang Baiao Biotechnology, Ltd were analyzed. 1999-2000 in Laiwu City.

RESULTSMost of the individuals belonged to rural population. In most cases the vaccination was done in spring and autumn. Persons between ages 18-40 years dominated the series. The side effects of the vaccination were seen in 1.71% in which local reaction was 1.35%, general reaction 0.30%, abnormal reaction 0.06% and the lowest reaction rate was seen in the group between ages 7-17.

CONCLUSIONSThe vaccine was quite effective. Both antibody positive ratio and GMT level of antibody were high.

Adolescent ; Adult ; Aged ; Child ; Female ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; prevention & control ; Humans ; Male ; Middle Aged ; Vaccination ; Vaccines, Inactivated ; Viral Vaccines

OBJECTIVETo understand the molecular mechanisms of hantavirus assembly and maturation by stably expressing the intracellular antibodies to hantavirus glycoprotein G1 and G2 in endoplasmic reticulum (ER) and cytoplasm (Cyto) of Vero E6 cell.

METHODSThe genes of VH and VL of antibodies against glycoprotein of hantavirus were amplified by PCR and cloned into pOPE 101-215 (Yol) vector. The G1 and G2 proteins specific ScFv genes were first expressed in E.coli and the function and binding properties were identified. The gene of ScFv were further inserted into intracellular expression vectyrs pEF/ myc/ ER and pEF/ myc/ CYTO vector and transfected Vero E6 cell. The clonal cell line which stabl expresses ScFv were isolated under the pressure of G418.

RESULTSThe ScFv genes of hantavirus G1 and G2 specific antibodies were successfully expressed in subcellular compartment ER and Cyto of Vero E6 cells and specifically targeted G1 and G2 protein after virus infection of the cells.

CONCLUSIONSThe recombination of intrabody to Hantann virus glycoprotein was constructed successfully, and it may provide basic material for the studying antiviral gene therapy and the molecular mechanism of viral replication and infection.

Animals ; Antibodies, Viral ; biosynthesis ; genetics ; immunology ; Cercopithecus aethiops ; Cloning, Molecular ; Endoplasmic Reticulum ; virology ; Glycoproteins ; biosynthesis ; genetics ; immunology ; Hantaan virus ; chemistry ; genetics ; immunology ; Humans ; Polymerase Chain Reaction ; Transfection ; Vero Cells ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen.

METHODSAn immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study.

RESULTSThe results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA.

CONCLUSIONThe results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level.

Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; analysis ; Early Diagnosis ; Female ; Fluorescent Antibody Technique, Indirect ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; immunology ; Humans ; Immunoblotting ; methods ; Immunoglobulin G ; immunology ; Male ; Rabbits ; Rats ; Sensitivity and Specificity

OBJECTIVETo identify and characterize the epitope associated with the virus attachment protein (VAP) of hantaan virus.

METHODSThe monoclonal antibody 3G1 was used as the ligand to biospan from a phage-displayed 12-amino acid peptide library, then the positive phage clones were chosen and sequenced. The amino acid sequences of them were compared with that of hantaan virus G2 in homology. The characteristics of positive phage were studied by IFA and ELISA. A decapeptide combining to cell membrane was observed under laser scanning confocal microscope (LSCM).

RESULTSThe conservative motif PX(1-2) HX(0-2) H displaying on positive clones shared homologous amino acid sequence with G2 96YPWHTAKCHY105.

CONCLUSIONG2 96YPWHTAKCHY105 might play some roles in virus binding to host cell, and might be a possible key epitope of hantaan virus VAP.

Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; immunology ; metabolism ; Hantaan virus ; immunology ; Microscopy, Confocal ; Oligopeptides ; immunology ; metabolism ; Peptide Library ; Vero Cells ; Viral Proteins ; immunology ; metabolism