• 대한바이러스학회지
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• 제29권1호
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• pp.23-31
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• 1999

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 human 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.

• KSBB Journal
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• 제13권6호
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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.

• 센서학회지
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• 제32권1호
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• pp.39-43
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• 2023

Microscale thermophysical property measurements of liquids have been developed considering the increasing interest in the thermal management of cooling systems and energy storage/transportation systems. To accurately predict the heat transfer performance, information on the thermal conductivity, heat capacity, and density is required. However, a simultaneous analysis of the thermophysical properties of small-volume liquids has rarely been considered. Recently, we proposed a new methodology to simultaneously analyze the aforementioned three intrinsic properties using heater-integrated fluidic resonators (HFRs) in an atmospheric pressure environment comprising a microchannel, resistive heater/thermometer, and mechanical resonator. Typically, the thermal conductivity and volumetric heat capacity are measured based on a temperature response resulting from heating using a resistive thermometer, and the specific heat capacity can be obtained from the volumetric heat capacity by using a resonance densitometer. In this study, we analyze methods to improve the thermophysical property measurement performance using HFRs, focusing on the effect of the ambience around the sensor. The analytical method is validated using a numerical analysis, whose results agree well with preliminary experimental results. In a vacuum environment, the thermal conductivity measurement performance is enhanced, except for the thermal conductivity range of most gases, and the sensitivity of the specific heat capacity measurement is enhanced owing to an increase in the time constant.

• 대한바이러스학회지
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• 제26권1호
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• pp.77-90
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• 1996

Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

• 대한바이러스학회지
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• 제28권3호
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• pp.275-285
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• 1998

Based on the geographic range and distribution of its rodent reservoir host, the European common vole (Microtus arvalis), Tula virus is likely to be widespread throughout Eurasia. Tula virus-infected voles have been captured in Central Russia, Austria, Czech and Slovak Republics, and the former Yugoslavia. Although serologic evidence for Hantaan (HTN) or Seoul (SEO) virus infection can be found in the vast majority of the more than 300 cases of hemorrhagic fever with renal syndrome (HFRS) occurring annually in Korea, approximately 4% of Korean patients with HFRS show a more than 4-fold higher antibody titer to Puumala (PUU) virus than to HTN or SEO virus by double-sandwich IgM ELISA, suggesting the existence of pathogenic Puumala-related hantaviruses in Korea. To further define the geographic distribution and genetic diversity of Tula virus in Eurasia and to investigate the existence of previously unrecognized Microtus-borne hantavirus in Korea, arvicolid rodents were captured in Lodz, Poland in 1995 and in Yunchon-kun, Kyungki-do during April to May, 1998. In addition, sera from 18 Korean HFRS patients who showed higher (or the same) antibody titer to Tula virus than HTN and SEO viruses were examined for hantavirus RNA by RT-PCR. Hantaviral sequences were not detected in any of the 18 patients or in 35 reed voles (Microtus fortis) in Korea. Alignment and comparison of a 208-nucleotide region of the S segment, amplified from lung tissues of two hantavirus-seropositive Marvalis captured in Poland, revealed $80.8{\sim}83.2%$ sequence similarity, respectively, with Tula virus strains from Central Russia and the Czech and Slovak Republics. Phylogenetic analysis indicated that the newfound Tula virus strains from Poland were closely related to other Tula hantaviruses from Eurasia.

• 대한미생물학회지
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• 제20권1호
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• pp.115-125
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• 1985

Hemorrhagic fever with renal syndrome (HFRS) is a debilitating disease of humans caused by Hantaan virus (HV), the prototype member of a newly proposed genus of Bunyaviridae. Studies of HV pathogenesis have been limited by the absence of a well defined model for a virus-induced disease state. In an attempt to devise a model for HV pathogenesis in laboratory rodents, newborn outbred suckling ICR mice were shown to be uniformly susceptible to lethal infection with non- mouse adapted HV by intracerebral (IC), intraperitoneal (IP), intramuscular (IM), and subcutaneous (SC) inoculation routes. Clinical coures, mean time to death, and fatal outcome were age-dependent. With an inoculum of 10 $LD_{50}$, mortality was 100% in mice infected within 72h of birth, but declined to 50% by 7 days. By 2-2.5 weeks, animals developed complete resistance to clinical disease. Virus was consistently detected in serum by day 6 post-infection in IC- and IP- inoculated animals, and reached peak levels of $10^5\;PFU/ml$ by day 8 Mice infected IM and SC showed delays in onset of viremia, but achieved similar titers. Immunofluorescent antibody appeared by 17-18 days, and neutralizing antibody by 15 days, in all experimental groups. Two of 8 inbred mouse strains were identified as resistant to clinical disease : SJL/J and A/J. Manipulation of this model will allow investigation of natural rodent pathogenesis with HV, as well as offer insight into disease mechanisms and therapy of HFRS.

• 대한바이러스학회지
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• 제29권3호
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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.

• 대한바이러스학회지
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• 제28권4호
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• pp.327-335
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• 1998

Hantaan (HTN) and Seoul (SEO) viruses, murid rodent-borne hantaviruses, are known to causes hemorrhagic fever with renal syndrome (HFRS) in Korea. To determine the genomic diversity and molecular phylogeny of HTN and SEO viruses found in Korea, we amplified for part of M and S genomic segments of hantaviruses from sera of HFRS patients and lung tissues of hantavirus seropositive striped-field mice. Both M and S segment of 16 HTN and 2 SEO viruses were amplified by nested reverse transcription-polymerase chain reaction. Based on 324 nucleotides in the M genomic segment, the HTN and SEO strains showed $93.8{\sim}100%$ and $99.1{\sim}99.4%$ homologies, respectively. Similarly, based on 230 nucleotides in the S genomic segment, HTN and SEO strains showed $90.9{\sim}100%$ and 100% homologies, respectively. Phylogenetic analysis of M and S segments indicated that HTN strains could be divided into at least two main groups in M and S trees and the sequence differences detected among the Sand M genomic segments of HTN viruses are consistent with reassortment having taken place between HTN virus strains.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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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.

• 대한바이러스학회지
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• 제27권2호
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• pp.177-183
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• 1997

Multiple species of muridae and arvicolidae rodents serve as the natural reserviors of hantaviruses. Hantaviruses are distributed in rodent populations world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Serologic diagnosis of infection, using hantaviral antigen, indicates that hantaviruses are widey distributed in wild rodents. This study was designed to intended the hantavirus infection among wild rodents captured in Kyebang mountain, Kangwon-do in Korea. A total of 216 wild rodents in 3 species were trapped in July and September in 1995. Serological evidence for hantaviruses infection were tested against five hantavirus antigens by indirect immunofluorescent antibody technique (IFA). Among 100 Eothenomys regulus, 78 Apodemus peninsulae and 38 Apodemus agrarius; 12 C. regulus, 15 A. peninsulae and 6 A. agrarius were IF antibody positive against hantaviruses. This data suggest that Eothenomys regulus and Apodemus peninsulae would be a natural reservoir of hantaviruses.