• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.53-58
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• 2002

To better understand the reservoir host range and distribution of hantaviruses in small mammal populations in Korea, a serological survey was conducted on 1,375 wild rodents and 62 insectivores captured in seven provinces during the six-year period, 1995 to 2000. As determined by the indirect immunofluorescent antibody (IFA) test, 90 ($13.1\%$) of 685 Apodemus agrarius, 47 ($13.6\%$) of 345 Apodemus peninsulae, and 4 ($6.5\%$) of 62 Crocidura laciura were seropositive against the Hantaan virus, while 38 ($13.5\%$) of 282 Eothenomys regulus were seropositive against the Puumala virus. Serological evidence for hantavirus infection was not found in 50 Microtus fortis, six Micromys minutus, six Mus musculus, and one Cricetulus triton. Our serological data indicate that hemorrhagic fever with renal syndrome (HFRS)-related hantaviruses are widely distributed in indigenous rodents in Korea. Particularly noteworthy was the high seropositivity rates among Apodemus peninsulae and Eothenomys regulus captured in certain mountainous regions, suggesting that HFRS may be under-reported among nearby residents or among individuals who might visit such areas for recreational or occupational purposes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.289-293
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• 2004

Hantaviruses are found worldwide and are known to cause two serious and often fatal human disease: hemorrhagic fever with renal syndrome(HFRS) and hantavirus pulmonary syndrome(HPS). The typical clinical prodrome consists of fever, chills, myalgia, headache, and gastrointestinal symptoms. Treatment usually involves maintenance of fluids, blood pressure, ventilation and electrolytes. We report a patient who had multisystem inflammatory symptom with Hantaan virus antibody positive accompanied by mild fever and myalgia. This case was diagnosed as HFRS. This patient was treated by Galgun hegi-tang. As a result of this treatment, symptoms were markedly improved.

• The Journal of Korean Society of Virology
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• v.28 no.2
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• pp.157-164
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• 1998

Various hantaviruses were isolated from HFRS patients and various rodent species, in many parts of the world. Bandicotas were captured at Yogyakarta, east region of Sumatura island, Indonesia; and 4 rodents species including Bandicotas were captured at Chiang Rai in Thailand during 1995. Sera were collected from captured bandicotas and other rodent spicies were screened for antibody test against Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Sin Nombre (SN) viruses by immunofluoresence antibody assay (IFA). Hantavirus antigen in lung tissues were tested by IFA. Among 55 captured Bandicota indica in Indonesia, 14 (25.5%) were antibody positive against HTN, SEO, PUU and SN virus. Hantavirus antigen were detected from 5 (9.0%) out of 55 lungs tested. Among 34 captured Bandicota indica in Thailand, 9 (26.5%) were antibody positive against HTN, SEO, PUU and SN virus. Among 34 lungs tissues of Bandicota indica examined, 3 (8.8%) were antigen positive. In other rodent species, antibody positive against Hantaviruses of Rattus rattus, Rattus losea and Mus cervicolor were 4/62 (6.5%), 5/25 (20%), 1/1 (100%), respectively. But no one has antigen in their lung tissues. Antigen positive lungs suspension were inoculated into vero E6 cells for virus isolation and 4 viruses were isolated from Indonesian Badicota and 3 viruses from Thailand.

• The Journal of Korean Society of Virology
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• v.29 no.2
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• pp.55-64
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• 1999

In Yugoslavia, hemorrhagic fever with renal syndrome (HFRS) is one of the important national health problem, but no vaccine has been used to prevent HFRS. Since first HFRS case in 1952, sporadic cases of HFRS occurred every year and over 4,000 registered cases with $1{\sim}16%$ mortality so far. We performed a prospective, randomized double-blind placebo-controlled trial to evaluate the effectiveness of $Hantavax^{TM}$ against HFRS in 3,900 healthy adults living in the endemic areas of Yugoslavia. 1,900 people were given 0.5 ml of Hantavax subcutaneously twice at one month interval and a booster shot at one year after. For controls other 2,000 healthy people were given 0.5 ml of physiolosical saline as a placebo. We investigated HFRS cases in both the vaccinated and nonvaccinated groups by monitoring the program for patient registration in the areas from 1996 to 1998, and the effect of vaccine was analyzed epidemiologically. No confirmed case of HFRS was observed among 1,900 Hantavax vaccinees, while 20 confirmed cases were observed among 2,000 nonvaccinated control group. There were no remarkable side effects among the vaccinees either locally or in general after inoculation of the vaccine. The Hantavax vaccine showed statistically significant protective efficacy against HFRS among Yugoslavian people.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1017-1025
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• 2006

Hantaviruses possess three RNA segments of negative sense. Co-infection of closely related hantaviruses may result in generation of a progeny virus with genomic polyploidy, containing a partial or complete set of genome originated from more than one parental virus. To characterize the formation of viral genomic polyploidy, cultured Vero-E6 cells were co-infected with two closely related hantaviruses, Hantaan and Maaji, and the progeny viruses examined. The genotype of plaque-purified viruses was analyzed by a virus-specific RT-PCR. Seventy percent (67/96) of the progeny virus was categorized as Hantaan and 3.3% (2/96) was classified as Maaji, whereas 20% (21/96) was considered polyploidy as they contained both types of the S RNA segment. Most of the polyploidy progeny viruses were unstable and gave rise to either one of the parental viruses or a reassortant after several rounds of plaque purification. No recombination between the heterologous pair of S RNA was observed for those polyploid viruses during three consecutive plaque-to-plaque passages. These data suggest that the viral polyploidy formation constitutes a primary mechanism underlying the generation of a newly emerged hantavirus.

• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.49-57
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• 1997

We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

• KSBB Journal
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• v.13 no.6
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• pp.662-668
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• 1998

Viruses belonging to the Hantavirus genus cause two acute severe illness in humans, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome(HPS). Among them, Hantaan virus is one of the most important viruses causing HFRS. Recombinant expression vectors, pKK-NP and pET-NP, with Hantaan viral nucleocapsid gene were constructed, and used to transform Eschericia coli BL21(DE3). Stability of the vectors in the host strain, and effects of some environmental conditions on the expression of nucleocapsid gene were studied. Expression vector, pKK-NP, was very unstable, and the expression level of nucleocapsid gene was very low compared to that of pET-NP. BL21(pET-NP) produced about 100 mg of N protein per liter of culture broth. Induction time did not show any significant difference on the expression level of nucleocapsid gen and cell growth. BL21(pET-NP) culture at 35$^{\circ}C$ showed a little higher expression level than at 30$^{\circ}C$ during growth phase, but reached to the same level at stationary phase. Total expression level was proportional to supplemented glucose concentration of media up to 0.5% along with cell growth, but expression level per unit cell mass was inversely proportional to glucose concentration and maximal when glucose was not supplemented at all.

• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.39-47
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• 1997

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 (SED), 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 occuring 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 $27\;{\mu}g/ml$, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is less 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.

• The Journal of Korean Society of Virology
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• v.29 no.3
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• pp.155-163
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• 1999

Hantaan virus (HTNV), the etiologic agent of hemorrhagic fever with renal syndrome (HFRS), belongs to the genus Hantavirus, and has three single negative stranded 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 L segment of Howang strain shows 93%, 74%, 66%, 65% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus $H{\ddot{a}}lln{\ddot{a}}s$ 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 $H{\ddot{a}}lln{\ddot{a}}s$ B1 and Sin Nombre virus, respectively.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.67-76
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• 1996

The methods that make Hantavirus grow consist of inoculation into the experimental animals and cultured cells. The cultured cells, such as Vero-E6 and A549 cells, have been usually used for isolation of the virus and the animals, such as mice and rats, are used for large scale preparation of the virus so far. Furthermore, the cell can be used to maintain the virus and assay the infectivity and the animals can be used for the experiment of viral pathogenicity and challenge for assessment of vaccine. Apodemus mice, the own natural host of the virus, has been used for challenge test of Hantaan virus. However it has been pointed out to difficult handling and breeding the animal in laboratory. Therefore, we attempted to establish a new animal model for challenge test at the time of isolation of Maaji virus which is a new hantavirus similar but distinct to Hantaan virus. In suckling hamster, the titer of Maaji virus and the lethality to mice of the virus were increased gradually in the titer and lethality through passage by intracerebral (IC) inoculation. We tried to re-adapt this brain virus to lung of weanling hamster. The brain passaged virus was inoculated into weanling hamster intramuscularly. Again, the titer of the virus in lung was also increased by continuous passage of this virus. This facts could regarded as adaptation to new environment in which the virus proliferates. To identity the virus passaged in hamster with Maaji virus, both of the virus passaged in hamster brain and lung were compared with Maaji virus (MAA-I) and Hantaan virus (HTN 76-118) by means of restriction fragment length polymorphism (RFLP) and slingle strand conformation polymophism (SSCP). As a result, we conclude that Maaji virus could be adapted successfully to weanling hamster through this passage strategy. Utilizing this adapted Maaji virus strain, hamster model is able to be used for challenge test in hantaviral vaccinology and further experiments utilizing hamster system as a rather available and convenient lab animal are expected.