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

In order to detect the nucleic acid of Puumala hantavirus, RNA was extracted from lungs of bank voles captured in Northeast China, and partial S and M genome segments of Puumala virus were amplified by RT-PCR and sequenced. Phylogenetic analysis suggested that Chinese Puumala virus had diverged from the common node of PUUV, with accumulating nucleotide substitutions and formed a distinct lineage from other Puumala viruses. Newly found Puumala virus was most closely related to the Kamiiso-8Cr-95 and Tobetsu-60Cr-93 strains which came from Japan and the muju strains which came from South Korea. By analysis of S and M genome segments of Puumala virus, we deduced a new Puumala virus subtype did exist in Northeast China.
Animals ; Base Sequence ; Cercopithecus aethiops ; China ; Evolution, Molecular ; Genome, Viral ; Hantavirus ; genetics ; Molecular Sequence Data ; Phylogeny ; Puumala virus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Rodentia ; virology ; Sequence Analysis, DNA ; Vero Cells

Apodemus agrarius, which accounts for three-fourths of the wild rodents, mainly inhabits in cultivated fields of Korea. Apodemus peninsulae and Eothenomys regulus are the second and third dominant species, respectively. Soochong virus (SOOV) from A. peninsulae and Puumala-related Muju virus (MUJV) from E. regulus were isolated in 1997 and 1998 in Korea, respectively. But serological characterizations of SOOV and MUJV were not identified clearly. Thus, in order to determine the serotypic classification, simultaneous cross-indirect immunofluorescent antibody (IFA) assay and cross-plaque reduction neutralization (PRN) test against four different hantaviruses were conducted with sera from 17 A. agrarius, 19 A. peninsulae, and 8 E. regulus strains. IFA titers of sera from A. agrarius and A. peninsulae were the highest to Hantaan virus (HTNV) and SOOV, respectively. However, most sera showed similar IFA titers to Seoul virus (SEOV). Therefore it was difficult to do serotyping using the sera from A. agrarius and A. peninsulae by IFA. In case of sera of E. regulus, IFA titers to Puumala virus (PUUV) were higher than HTNV, SOOV and SEOV. Cross-PRN result of A. agrarius to HTNV, SOOV, SEOV and PUUV was 6,890, 5,120, 110 and 30, respectively. In case of A. peninsulae, the mean PRN titer was the highest to SOOV (1:6,820) and those to HTNV, SEOV and PUUV were 1,580, 100 and 30, respectively. The mean PRN titers of E. regulus to HTNV, SOOV, SEOV and PUUV were 70, 10, 80 and 640. SOOV and MUJV could be distinguished from HTNV and SEOV by cross-PRNT. These results demonstrate that SOOV and MUJV could be classified as new serotype of hantavirus.
Animals ; Classification ; Hantaan virus ; Hantavirus* ; Korea ; Murinae ; Puumala virus ; Rodentia ; Seoul virus ; Serotyping

BACKGROUNDTo confirm if Puumala like viruses exist in China.

METHODSRNA was extracted from lungs of bank voles captured in the Northeast China, partial S segments genome of Puumala viruses were amplified and sequenced.

RESULTS926 bp cDNA of S segments of Puumala like virus was amplified and sequenced. The phylogenetic analysis revealed that the Puumala like viruses found in China were most close to that found in Far East region of Russia.

CONCLUSIONSPuumala like virus does exist in Northeast China, and the nucleotides sequence of the viruses have high homolog to Puumala viruses found in Russia.

Animals ; China ; DNA, Viral ; analysis ; Hemorrhagic Fever with Renal Syndrome ; virology ; Lung ; virology ; Mice ; Phylogeny ; Puumala virus ; isolation & purification ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

Hantaan virus (HTNV), the etiologic agent of hemorrhagic fever with renal syndrome (HFRS), belongs to the genus Hantavirus, and has three single negative straded RNA genome segments. HTNV strain Howang isolated from the blood of severe case of Korean HFRS is more virulent than HTNV 76/118 and the M and S genome segments' nucleotide sequence of Howang strain showed 93.5% and 94% homology to each segment of HTNV 76/118. We have obtained 6533 nucleotides long sequence of the L genome segment of Howang strain using reverse transcriptase in conjunction with PCR amplification and compared to other hantaviruses. The messenger sense of the L segment contains one long single long open reading frame of 2151 amino acids, which encodes a deduced RNA dependent RNA polymerase of 246.4 kDa caculated molecular weight protein. The nucleotide sequence of the Lsegment of Howang strain shows 93%, 74%, 66%,65% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus Hallnas B1 and Sin Nombre virus, respectively. The amino acid sequence of the L segment of Howang strain shows 99%, 85%, 68%, 68% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus Hallnas B1 and Sin Nombre virus, respectively.
Amino Acid Sequence ; Amino Acids ; Base Sequence ; Genome ; Hantaan virus ; Hantavirus ; Hemorrhagic Fever with Renal Syndrome ; Molecular Weight ; Nucleotides ; Open Reading Frames ; Polymerase Chain Reaction ; Puumala virus ; RNA ; RNA Replicase ; RNA-Directed DNA Polymerase ; Seoul virus ; Sin Nombre virus

Hantavirus is a genus of the Bunyaviridae family causing two serious diseases, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Puumala virus is a member of hantavirus originally found in Europe, and its natural reservoir is Clethrionomys glareolus. It is also associated with the hurnan disease nephropathia epidemica, a milder form of HFRS. To identify the hantaviruses in bats, bats were collected from Jeong-Sun, Won-Joo, Chung-Ju and Hwa-Cheon area in Korea, and nested RT-PCR was performed with serotype specific primer from M segment. Interestingly, Puumala virus was detected in bats (Rhinolophus ferrum-equinum) only from Won-Joo. The 327 bp nested RT-PCR product, was sequenced. The sequence database search indicates that the sequence is homologous to the published sequence of Puumala viruses. The sequence similarities were ranged from 71% to 97%. The highest sequence similarity was 97% with Puumala virus Vranicam strain, and the lowest was 71% with Puumala virus K27 isolate. Puumala virus Vranicam strain was isolated from a bank vole (Clethrionomys glareolus) in Bosnia-Hercegovina. Puumala virus K27 was isolated from human in Russia. This analysis confirms that bats (Rhinolophus ferrum-equinum) in Korea are natural reservoir of Puumala virus.
Arvicolinae ; Base Sequence ; Bunyaviridae ; Chiroptera* ; Chungcheongbuk-do ; Europe ; Hantavirus ; Hantavirus Pulmonary Syndrome ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Korea* ; Puumala virus* ; Russia ; Sequence Analysis*

Hantaviruses are distributed in rodent population world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Va.ictus species of Family Muridae and Arvicolidae serve as the natural reservoirs of hantaviruses. Hantaan virus, Seoul virus, Puumala virus, Prospect Hll virus, Sin Hombre virus and New York virus are members of genus Hantavirus and isolated from lungs of A. agrarius, C glareolus, M. pennsylvanicus, P. maniculatus and P. leucopus respectively. This experiment was intended to find the distribution of hantavirus infection among wild rodents and isolate the hantavirus from lung tissue of seropositve Apodemus peninsulae, and compared the nucleotide and amino acid sequences with prototype of hantaan virus 76-118 strain. Hantaviral sequences were amplified from lung tissues of A. peninsulae by reverse-transcriptase polymerase chain reaction. Alignment and comparison of the 324 nucleotide of G2 region of M-genomic segment diverged 4.6% and 0% at the nucleotide and amino acid levels, and complete N protein-coding region of S-genomic segment diverged 3.7% and 1.4% nucleotide and amino acid levels, respectively. This is the report to spill-over on the hantaan virus from A. peninsulae to A. peninsulae in Korea.
Amino Acid Sequence ; Animals ; Gyeonggi-do* ; Hantaan virus ; Hantavirus Infections ; Hantavirus* ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Korea ; Lung ; Muridae ; Murinae* ; Polymerase Chain Reaction ; Puumala virus ; Rodentia ; Seoul virus

Eleven hantavirus isolates were obtained by innoculation of viremic blood, urine, or autopsy tissue specimens from ten HFRS patients, and sera were obtained from five patients with HFRS. The disease was diagnosed by clinical manifestations and indirect immunofluorescent antibody technique. We obtained 6 hantaviruses from gene bank. So, we analyzed 22 hantavirus samples to elucidate the genetic diversity. The hantaviral RNAs were extracted and 365 base-pair complementary DNAs of M segment were obtained by reverse transcriptase polymerase chain reaction (RT-PCR) and 326 base-pair by nested PCR. The nucleotide sequences of amplified cDNA fragments were determined by the direct sequencing method using automatic DNA sequence analyzer. We got full M segment sequences of 28 reported hantaviruses with medline searching, and aligned them with our 22 samples, and the phylogenetic analysis for nucleotide and amino acid sequences were done by the Clustal method. The nucleotide and amino acid sequences of Hantaan virus 17 samples showed high (above 90%) homology with 76-118 strain, but 2 samples showed significant differences with 76-118 strain and with other 17 samples. The 3 Seoul virus samples showed high intraspecies differences in 1 sample, and showed singnificant differences with SR-11 strain. In phyogenetic tree analysis, Puumala virus and Hantavirus pulmonary syndrome viruses showed high homology, but Hantaan and Seoul viruses showed significant genetic diversity among strains. In conclusion, hantaviruses isolated from HFRS patients showed genetic diversity compared with those isolated from rodent hosts.
Amino Acid Sequence ; Autopsy ; Base Sequence* ; DNA, Complementary ; Genetic Variation ; Hantaan virus ; Hantavirus Pulmonary Syndrome ; Hantavirus* ; Hemorrhagic Fever with Renal Syndrome* ; Humans ; Korea* ; Polymerase Chain Reaction ; Puumala virus ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; Rodentia ; Seoul virus

Hantavirus is a genus of the Bunyaviridae family consisting following serotype groups: Hantaan, Seoul, Puumala, Prospect Hill, Thailand, Belgrade, Thotta palayam, Sin Hombre. Most of Hantavirus group have been associated with many clinically similar disease known collectively as hemorrhagic fever with renal syndrome (HFRS). Hantaan virus is the prototype of the genus hantavirus, originally isolated from Apodemus agrarius. Bat was found as a natural host for Hantaan virus in Lee's lab for the first time. Then, Hantaan-like virus was isolated Hantaan-like virus from bat. To identify hantaviruses that are present in Korea among bats, bats were collected from Jeong-Sun, Won-Joo, Chung-Ju and Hwa-Cheon area, RNA was isolated from lung and serum. RT-PCR was performed with a universal primer from M segment. Nested RT-PCR was carried out to differentiate Hantaan, Seoul and Puumala virus using serotype specific primers. As we expected, Hantaan viruses were detected in bats and Seoul virus was not detected. Interestingly, Puumala viruses were also detected in bats from won-Ju, but not in other areas. Puumala virus is originally isolated from Clethrinomys glareolus, and cause light HFRS. Recently, Paradoxomis webbiana, a wild bird turn out to be a reservoir for Puumala virus in Korea. These data indicate that bat is a new natural reservoir of Puumala virus.
Animals ; Birds ; Bunyaviridae ; Chiroptera* ; Chungcheongbuk-do ; Gangwon-do ; Hantaan virus* ; Hantavirus ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Korea* ; Lung ; Murinae ; Puumala virus ; RNA ; Seoul ; Seoul virus ; Thailand

A large number of viruses belonging to Genus Hantavirus in Family Bunyaviridae are etiologic agents for hemorrhagic fever with renal syndrome (HFRS), or hantavirus pulmonary syndrome (HPS). Hantaan (HTN), Seoul (SEO), Belgrade (BEL), Puumala (PUU) serotype viruses are well known causative agents for HFRS in Eurasian continent. Among those viruses Hantaan and Seoul serotypes are well known to cause HFRS in Korea, but there are some sporadic incidence by other than Hantaan or Seoul viruses. Recently we have developed the combined Hantaan-Puumala virus vaccine to prevent world-wide occurring HFRS. This combined vaccine is formalin inactivated, suckling mouse and suckling hamster brain extracts for Hantaan and Puumala viruses, respectively. Protein contents of this purified candidate vaccine is 27 microgram/ml, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is legs than 0.1 ng/ml. Thirty hamsters were given twice at one month interval intra-muscularly and bled on 30 days after each vaccination from retro-orbital sinus vein. Antibody titers were tested against 5 major serotype viruses, Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses by IFA and PRNT. The mean IF antibody titers on 30 days after primary shot were 78.4, 68.8, 68.8, 37.9, and 15.6; mean neutralizing antibody titers were 65.4, 12, 6.1, 65.6 and 0.5 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively. The mean IF antibody titers on 30 days after booster shot were 686.9, 567.5, 550.4, 516.3, and 430.9; and neutralizing antibody titers were 710.8, 41.9, 24.3, 409.9, and 1.6 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively.
Animals ; Antibodies, Neutralizing ; Brain ; Bunyaviridae ; Cricetinae* ; Encephalomyelitis, Autoimmune, Experimental ; Enzyme-Linked Immunosorbent Assay ; Formaldehyde ; Hantavirus ; Hantavirus Pulmonary Syndrome ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Incidence ; Korea ; Leg ; Mice ; Myelin Basic Protein ; Puumala virus ; Seoul ; Seoul virus ; Sin Nombre virus ; Vaccination ; Veins