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

Animals ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; virology ; Molecular Epidemiology ; Puumala virus ; genetics

OBJECTIVETo understand the rate of viral carrying status among rodents as well as genotypes and distribution of Hantaviruses (HV) isolated in Hunan province.

METHODSWith DFA, the HV antigen in lung tissues of rodents was detected. The total viral RNA was extracted from the lung tissues of the HV infected rats and amplified with reverse transcrition-polymerase chain reaction (RT-PCR), using the HV genotype specific primers. The amplified genes were then sequenced and subjected to genotyping and homologic analysis.

RESULTSThe average density of rodents was 3.15% and the virus carrying rate among rodents was 1.31%. Data from genotype analysis showed that the HV isolated from seven lung specimens taken from Rattus norvgicus, Apodemus agraius, Mus musculus, Rattus flavipectus among indoor rodents in Shaodong and Liuyang belonged to HV type II (SEOV), and one isolated from Apodemus agraius in Shaungfen belonged to HV type I (HTNV) among outdoor rodents. Six strains were sequenced successfully and the homology between six srains was 88.3%-100%. The homology of HN1, HN2, HN4, HN6 came from Liuyang and the HN7 and HN8 from Shaodong were both 100% while the homology between L99 and the strains from Liuyang and Shaodong were 94.4% and 88.3% respectively.

CONCLUSIONHV type II (SEOV) and the HV type I (HTNV) were all existed in Hunan province while SEOV was the main genotype.

Animals ; Genotype ; Hantavirus ; classification ; genetics ; Hemorrhagic Fever with Renal Syndrome ; virology ; Phylogeny ; RNA, Viral ; genetics ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVES: In HFRS, there is a varying degree of disseminated intravascular coagulation which was evident in the early phase of the illness. It is believed also that DIC would be the consequence, at least in part, of functional changes of endothelium resulting in kinin activation and clinical syndrome. This study investigated the role of adhesion molecule in the pathogenesis of Hantaan virus-related disease. METHODS: The expression of ICAM-1 antigen on the cell membrane of human umbilical vein endothelial cells was assessed by immunohistochemistry, and ICAM-1 mRNA in the endothelial cells was assessed by in situ hybridization after Hantaan virus infection (2.6 x 10(4) PFU/mL) with the time course. RESULTS: In immunohistochemistry, the number of ICAM-1 positive cells increased with time during the 12 or 24 hours after infection. 5 to 10% of HUVECs had been positive after 12-24 hours and the number of positive cells decreased abruptly after 24 hours. Hantaan antigen had been noticed after 12 hours focally on the HUVECs but continued to proliferate into day 7 post-infection when most of HUVECs were infected by Hantaan virus. In situ hybridization showed identical patterns of ICAM-1 mRNA expression after Hantaan virus infection. CONCLUSION: It implies that the Hantaan virus infection on HUVECs would express more ICAM-1 on their surface and implicated in the pathogenesis of early clinical syndrome of HFRS.
Cell Line ; Endothelium, Vascular/virology ; Endothelium, Vascular/immunology ; Gene Expression ; Hantaan Virus/pathogenicity* ; Hemorrhagic Fever with Renal Syndrome/immunology* ; Hemorrhagic Fever with Renal Syndrome/genetics ; Hemorrhagic Fever with Renal Syndrome/etiology ; Human ; Immunohistochemistry ; In Situ Hybridization ; Intercellular Adhesion Molecule-1/metabolism* ; Intercellular Adhesion Molecule-1/genetics* ; RNA, Messenger/metabolism ; RNA, Messenger/genetics

OBJECTIVETo investigate whether Hantavirus (HV) and Orientia tsutsugamushi ( OT) can naturally infect and coexist in their host and role.

METHODSBy field epidemiological study, Leptotrombidium scutellare (3829) was collected and separated from mice(166) in epidemic areas. The cells of mites separated from their host and role were cultured. PCR was used to detect HV-RNA and OT-DNA in the cell culture.

RESULTSIn 105 Apodemus agrarius, 3 HV-RNA positive, 2 OT-DNA positive and 2 coinfection with HV and OT were detected;in 41 Brown rattus, 2 HV-RNA positive, 1 OT-DNA positive and 1 co-infection with HV and OT were detected. From 15 mites co-infected with HV and OT, 2 strains of HV pathogen, 2 strains of OT pathogen were separated and 1 HV and OT pathogen in the same mite were separate.

CONCLUSIONThe study demonstrates that co-infection of HV and OT did simultaneously exist in wild Leptotrombidium scutellare. This theory has some significance to the epidemic and precaution of HV and OT.

Animals ; Disease Vectors ; Hantavirus ; genetics ; pathogenicity ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; Host-Parasite Interactions ; Orientia tsutsugamushi ; genetics ; pathogenicity ; Rats ; Scrub Typhus ; epidemiology ; Trombiculidae ; microbiology

OBJECTIVETo know the genotype and subtype of hantavirus (HV) which infected persons in Hebei province.

METHODSAccording to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients' sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package.

RESULTS17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, or= 14 days were 0. 17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7%-100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens' nucleotide homology was only 70.3%-72.7%, belonged to different type.

CONCLUSIONSEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and S1 was also existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability.

China ; Genotype ; Hantavirus ; classification ; genetics ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; prevention & control ; therapy ; virology ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

BACKGROUNDTo observe the features of serum specific IgA, IgG, IgM antibodies in the acute phase of hemorrhagic fever renal syndrome (HFRS).

METHODSThe nucleocapsid (NP) protein and glycoproteins (GP) of Hantavirus were expressed by recombinant baculovirus, and used as ELISA antigens to test 61 serial sera of 14 acute phase HFRS patients.

RESULTSSeoul like virus RNA were detected from 11 of 14 patients. An early and strong IgA, IgG and IgM antibody response to recombinant NP (rNP) was observed in almost all HFRS cases. The titers of antibody to rNP was apparently higher than that to Rgp. In the early stage, titer of IgG antibody elevated most drastically among all the three classes of antibodies to rNP, followed by IgM and IgA antibody responses. The elevation trend of IgM and IgA antibodies to rNP stayed nearly at the same level, but the IgA titers to rNP were apparently higher than that of IgM. Among the antibodies to rGP, IgA changed distinctly greater than IgG. The elevation trend of IgM could be found during first week after the onset, and the titers dropped gradually after the second week. IgM antibodies of one case who was viral RNA positive were not detected at early stage, but IgA titers were high. The only severe case of the 14 patients kept the lower IgA, IgG and IgM during the whole acute phase.

CONCLUSIONHFRS patients kept an early and strong humoral response to NP and GPs in acute phase of HFRS.IgA could be used together with IgM to improve the diagnostic accuracy.

Acute Disease ; Adult ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; Female ; Hantavirus ; genetics ; immunology ; Hemorrhagic Fever with Renal Syndrome ; immunology ; virology ; Humans ; Immunoglobulin Isotypes ; blood ; Male ; Viral Core Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

OBJECTIVETo study the molecular epidemiological characteristics of hantavirus seen during 2000-2003 in Qingdao region of Shandong province.

METHODSSera were collected from 64 patients with hemorrhagic fever with renal syndrome (HFRS) and viral RNA was extracted from the sera. HTN and SEO universal primers were designed as outer primers and HTN and SEO specific primers as inner primers. G1 gene region of M segment from hantavirus was amplified by using RT-nest-PCR for sequencing. The data of nucleotide sequences were analyzed by DNA star software.

RESULTSSix cases were positive by HTN specific primer of total cases (9%); 25 of 64 cases by SEO specific primer (39%); total positive rate was 48%. In general, SEO type was a prevalent type of hantavirus in Qingdao region. The variation of the nucleotide sequences among SEO viruses (nucleotide sequence divergence ranged from 0.3% approximately 8.9%) was lower than that among HTN type (nucleotide sequence divergence ranged from 2.6% approximately 11.2% ).

CONCLUSIONMajority of hantavirus found in Qingdao region belonged to SEO type and still a few strains belonged to HTN type. Most of the HTN viruses were detected in Jiaonan county.

China ; Genotype ; Hantavirus ; genetics ; Hemorrhagic Fever with Renal Syndrome ; blood ; virology ; Humans ; RNA, Viral ; blood ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo understand antibody responses to and RNA sequences of Hantavirus in patients with hemorrhagic fever renal syndrome (HFRS) in Yantai areas and to demonstrate the type of the prevalent viruses caused HFRS.

METHODSSerum specimens collected at acute and convalescent stages from 90 patients with HFRS and IgM and IgG antibodies against Hantavirus were detected with ELISA, and cross plaque reduction neutralizing tests were performed to detect neutralizing antibody. Viral RNA was extracted from the patients? sera by using Trizol method and nested PCR was utilized to amplify the specific segments of the viral cDNA and the products of the PCR were TA cloned and then the nucleotide sequences were determined.

RESULTSThe IgM antibody was positive in 82.2% (88/107) of the patients while the IgG antibody was positive in 85.7% (66/77) of the patients. Both the serologic and sequence analyses demonstrated that the epidemic of HFRS in Yantai areas was caused by mixed types of Hantavirus. The prevalent strains of Hantavirus had higher homology with the strains isolated in Korea than with those isolated previously in China.

CONCLUSIONSThe serologic and sequencing analyses indicated that the epidemic of HFRS in Yantai areas was caused by mixed types of Hantavirus dominated by type SEO.

Antibodies, Viral ; blood ; Base Sequence ; China ; DNA, Viral ; analysis ; Disease Reservoirs ; Hantaan virus ; classification ; genetics ; immunology ; Hemorrhagic Fever with Renal Syndrome ; virology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Molecular Sequence Data ; Sequence Analysis, DNA ; Serotyping

OBJECTIVETo compare the molecular characteristics of hantaviruses isolated from Shandong province by using PCR typing and nucleotide sequencing.

METHODSTotal cellular RNA was extracted from hantaviruses infected Vero E6 cells, viral cDNA was amplified by polymerase chain reaction. PCR products of partial M Segments of 4 strains of hantaviruses were sequenced. Cross neutralization tests were performed.

RESULTSFour strains of hantavirus isolated from Rottus in Shandong province were SEO like viruses. The homology between Shandong isolates and other SEO like viruses was high at both amino acid and nucleotide levels. The homology among those 4 strains of hantaviruses was 98%.

CONCLUSIONSThe SEO like hantaviruses were more conserved than other hantaviruses. The homology of SEO like hantaviruses isolated from Shandong province was as high as 98% at nucleotides level, though they were isolated at more than 10-year intervals.

Animals ; Base Sequence ; China ; DNA, Viral ; analysis ; Disease Reservoirs ; Genetic Variation ; Hantaan virus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever with Renal Syndrome ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid