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ABSTRACT: In Korea, there was a big outbreak of aseptic meningitis in 1993. Six clinical isolates of enterovirus were obtained from patients with aseptic meningitis and were identified as echovirus type 9 by serotyping with a pool of neutralizing antisera. For molecular characterization of the isolates, the nucleotide sequences of 5'-noncoding region (NCR), VP4, VP2, VP1, 2A and 2C regions of the isolates were compared with the corresponding regions of echovirus type 9 Hill and Barty strains. Unlike Hill strain, Barty strain contained a C-terminal extension to the capsid protein VP1 with an RGD (argnine-glycine-aspartic acid) motif. To determine whether similar structural features were present in our isolates, their nucleotide sequences including the VP1 region were analyzed. All isolates exhibited the VP1 extension with the RGD motif. We concluded the Korean isolates in the year of 1993 as the echovirus type 9 Barty strain although the isolates showed 15-20% nucleotide sequence differences in the several genomic regions.
Experimental and Molecular Medicine 07/1998; 30(2):101-7. · 2.48 Impact Factor
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ABSTRACT: In Korea, there was a big outbreak of Aseptic Meningitis due to enterovirus infection in 1993. Since virus isolation and neutralizing tests are too laborious and time-consuming for the detection of enterovirus from clinical specimen, we have developed a new molecular identification method for rapid subgrouping of isolates from patients with aseptic meningitis. For the rapid subgrouping of isolates, RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and RFLP (Restriction Fragment Length Polymorphism) assays were used. We have selected two oligonucleotide primers from the conserved 5'-UTR/VP2 and VP1 regions. A 652 bp (base pair) product was amplified from the 5'-UTR/VP2 region of reference viruses and the isolates. For the subgrouping of the isolates by RFLP assay, we have used 12 reference viruses (Echovirus, E6, E9, E11, E12, Coxsackievirus, CB1, CB3, CB4, CB5, Coxsackievirus, CA9, CA16, CA21, CA24), which are the common viral agents associated with aseptic meningitis. By using subgroup-specific restriction enzymes BsmAI, , HinP1I, and PleI, the isolates were classified into Echovirus subgroups. We have also shown that subgrouping of the isolates by RFLP assay based on the VP1 region is possible.
Molecules and Cells 07/1998; 8(3):330-5. · 2.18 Impact Factor
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Y M Park,
M Mizokami,
T Nakano,
J Y Choi,
K Cao,
B H Byun,
C H Cho, Y T Jung,
S Y Paik,
S K Yoon,
M Mukaide,
B S Kim
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ABSTRACT: GB virus C and hepatitis G virus (GBV-C/HGV) have been identified from the patients with acute or chronic liver diseases as possible agents of non-B, non-C hepatitis by two different groups, independently. To investigate whether GBV-C/HGV plays a role among Korean patients with liver diseases, GBV-C/HGV RNA were evaluated in 337 sera by the reverse transcription polymerase chain reaction (RT-PCR) using specific primers derived from 5'-noncoding region of GBV-C/HGV genome. GBV-C/HGV RNA was identified in 11/337 (3.3%). They consisted of 1/160 (0.6%) and 10/177 (3.3%) among the general population and patients with liver diseases, respectively (P < 0.01). Nucleotide sequences of all PCR amplicons were determined by the dideoxy chain termination method and analyzed by molecular evolutionary methods. The phylogenetic tree showed all sequences could be divided into three genotypes. These results indicate that: (1) GBV-C/HGV already exist in Korea; (2) GBV-C/HGV may play some role as an etiologic factor among the Korean patients with liver diseases; (3) GBV-C/HGV infection is rare among Korean general population; and (4) there are at least three different types of GBV-C/HGV in Korea.
Virus Research 05/1997; 48(2):185-92. · 2.94 Impact Factor
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ABSTRACT: A part of the 5'-noncoding region of echovirus type 9 isolates was sequenced, and an attempt was made for rapid virus detection in clinical samples obtained from 22 subjects hospitalized with aseptic meningitis. The sequence identity of 440-bp products amplified from the region by RT-PCR was 87.7% between the standard echovirus type 9(Hill strain) and the isolates. Specific IgM antibodies to Hill strain were positive in 45.5% by immunofluorescent antibody staining of virus-infected cells. A high detection rate of PCR products was observed in cerebrospinal fluids (CSFs; 54.5%) at admission, and in peripheral mononuclear cells (PMCs; 72.7%) at the end of hospitalization. Viral genomes were detectable for 2 days in serum samples, and for 6 days in PMC samples after onset of disease. When specific IgM antibody titers were less than 1:40, the amplification rate of viral genome from serum samples was 50.0%. These results indicate that the combination of specific IgM determination and viral genome amplification from CSFs will be a rapid and reliable method for early diagnosis.
Archives of Virology 02/1997; 142(4):853-60. · 2.11 Impact Factor
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ABSTRACT: Most colon cancers exhibiting microsatellite instability (MI), a mutator phenotype of mismatch repair failure, are associated with mutations of the transforming growth factor-beta receptor type II genes (TGF-beta RII). Of intestinal- and diffuse-type gastric carcinomas, the former have been thought to arise from intestinal metaplasia in which gastric mucosa resembles intestinal mucosa. To evaluate the preferential histological type of MI-associated mutations in the development of gastric carcinoma, mutations of TGF-beta RII, p53, and p16 were analyzed for the two types of primary gastric carcinomas showing MI. Of 50 primary gastric carcinomas, including 33 intestinal types and 17 diffuse types, 15 cases (30%) demonstrated MI at 1 or more of the 11 microsatellite markers tested. The 15 MI cases were classified into two groups, widespread MI and low-level MI, based on the number of markers exhibiting the instability. Eleven were widespread MIs, and the remaining four cases were low-level MIs. Ten of the 11 (91%) widespread MIs were of the intestinal type, and 1 case (9%) was of the diffuse type. Of the 11 widespread MIs, 10 cases (91%) demonstrated frameshift mutations within the polyadenylate tract of the TGF-beta RII. The frameshift mutation was rarely detected at p53 and p16 (1 of 11, 9%). In contrast, the four low-level MI cases had no frameshift mutations within the repeat sequences of TGF-beta RII, p53, and p16, but two of the four cases demonstrated base substitution mutations within p53. Our results suggest that mismatch repair failure can mutate the TGF-beta RII and may provide one of the pathways for the development of the intestinal-type gastric carcinoma in high-risk populations.
Cancer Research 11/1996; 56(20):4662-5. · 7.86 Impact Factor
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ABSTRACT: Genomic analysis of three Hantaan-like virus isolates from bats was performed. Cleavage patterns of reverse transcription (RT)-polymerase chain reaction (PCR) products and nucleotide sequences of G2 region of M RNA segment and N protein region of S RNA segment of the isolates were compared to that of Hantaan 76-118 strain. Genomic characteristics of the bat isolates were identical to that of Hantaan virus.
Acta virologica 10/1995; 39(4):231-233. · 0.68 Impact Factor
