• 대한미생물학회지
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• 제18권1호
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• pp.39-46
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• 1983

We collected three hundred thirty-five strains of Salmonella typhi isolated from human sources during the period January to December 1982 Korea. Most of them were from general hospitals and city health center, the remaining one hundred sixtyone strains were obtained from other 12 provincial health centers among 335 testing strains. We used ninety-nine Vi-phages as distributed from International Center for Enteric phage Typing(ICEPT) in London. We found nine phage types among 335 strains of Salmonella typhi in this study. Additionally, I+IV, degraded Vi-positive and Vi negative strains were presented. This study documented the occurrence of the new $B_2$ type in Korea for the first time. The present basic phage type formula in Korea appeared to be A, $B_2,\;D_1,\;D_2,\;D_4,\;D_6,\;D_8,\;D_{12},\;E_1,\;E_9,\;D_1,\;M_1$, 40 and plus I+IV and degraded vi strains(Table 2). The current phage types of Salmonella typhi isolated in Korea 1982 were A, $B_2,\;D_1,\;D_2,\;D_6,\;D_8,\;E_1,\;M_1$ and 46 $M_1$ type was widely distributed all over the country, and E type was next predominant. Antibiotic susceptibility test were performed by means of Kirby-Bauer disc diffution method using 12 kind of antibiotics such as Ampicillin, Carbenicillin, Cephalosporin, Chloramphenicol, Colistin, Gentamicin, Kanamycin, Nalidix acid, Neomycin, Polymyxin-B, Streptomycin and Tetracycline. The sensitivity pattern to antibiotics of Salmonelia typhi cultures were summarized.

• 약학회지
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• 제45권5호
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• pp.536-544
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• 2001

increase the viability of oral typhoid vaccine during the passage through the castro-intes-tidal tract, numerous attempts have been made including the vaccine coating. However problems such as high death rate during the coating process and its instability in the gastric juice still remain to be solved. In this study, the oral vaccine was made as the micro-enteric beads by adding Salmomella typhi Ty21a cells to sodium alginate solution and spraying onto calcium chloride solution (ionotropic relation method). The vaccine showed more than 90% of its original viability after treating it for 1 hour in the artificial gastric juice (37$^{\circ}C$, 300 rpm). The clearance rate of the Ty21a in the liver and spleen of the mice orally administrated with coated Ty21a was similar to that of the mice intraperitoneally administrated with uncoated Ty21a. The peripheral blood lymphocytes (PBL) isolated from the mice orally administered with this vaccine produced 15.5 fold higher specific IgA antibody titer than that from the control mice administerd with saline solution. furthermore, the mice treated with the coated Ty21a had higher survival rates (50~87%) than the control mice treated with saline solution (0~10%) in the intraperitoneal challenge test with wild type S. typhi Ty21a cells. These results suggest that the alginate-based coating technique is effective to protect live Ty21a from acidic environments, and produces better intestinal immune responses thereby providing a potentially excellent oral typhoid vaccine.

• Pediatric Infection and Vaccine
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• 제20권1호
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• pp.1-8
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• 2013

목적 : 장티푸스와 세균성 위장염의 주된 원인인 살모넬라균에 의한 감염증은 우리 주변에서 흔하게 관찰할 수 있는 질환이며 최근 장티푸스의 발생 수는 줄고 있으나 살모넬라 감염증은 꾸준히 보고되고 있다. 이에 본 연구에서는 최근 12년간 살모넬라증 환자의 임상적 양상과 배지에서 동정된 살모넬라 균주의 변화, 항생제 내성을 연구하기로 하였다. 방법 : 2000년 1월부터 2011년 12월까지 12년간 분당 차병원에 입원한 18세 이하의 환아를 대상으로 하였으며, 대상자의 일반적인 특성을 의무기록을 통하여 후향적으로 조사하여 환아의 임상양상을 비교 분석하였다. 결과 : 대상 환아 수는 총 53례였으며 환아의 연령별 분포를 보면 2세가 10례로 가장 많은 분포를 보였으며 중간값은 3세(최소 12일, 최대 18세)였다. 대상 환아 중 남아의 수는 33례(62.3%)이다. 계절별로는 여름(6,7,8월)이 21례(39%)로 가장 많은 빈도를 보였으며, 연도별로 비교하였을 경우 2001년이 11례로 가장 많았으며, 2003년에는 7례가 진단되었으나 2004년에는 2례로 급감하는 양상을 보였으며 2009년과 2010년에는 진단된 환아가 없었다. S. typhi가 배양된 환아는 모두 3명이었으며 모두 혈액에서 S. typhi가 배양된 균혈증을 보이고 있었다. 항생제에 대한 감수성 결과에서는 ampicillin에 대한 내성이 16례로 가장 높았으며 S. typhi는 항생제 감수성 결과 내성을 보이는 항생제가 없었다. 2000년 1월부터 2005년 12월까지와 2006년 1월부터 2011년 12월까지의 두 군으로 나누어 전 6년군과 후 6년군으로 나누었으며, 전 6년군의 환아는 40례였으며 후 6년군의 환아는 13례였다. 연도에 따른 ampicillin에 대한 내성율은 후 6년군에서 더 증가한 소견을 보이고 있으나 통계학적인 의미는 없었으며(25% vs. 38.5%, P=0.349) 연도에 따른 혈청군별 항생제 내성율의 분포상 전 6년군에서는 D군이 가장 많았고, 후 6년군에서는 C군과 D군이 많았다. 결론 : 본 연구 결과 살모넬라 감염증은 2000년대 초반에 많았으나 2004년부터 급감하는 소견을 보이고 있으며 연도별로 본 혈청형과 항생제 내성의 경향에는 뚜렷한 변화가 관찰되지 않았다.

• 대한미생물학회지
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• 제20권1호
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• pp.91-102
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• 1985

The advantages of enzyme-linked immunosorbent assay(ELISA) are its senstivity and simplicity in detecting IgG, IgM and IgA antibody. To apply ELISA to diagnosis of typhoid fever, antigen such as lipopolysaccharide of Salmonella typhi or killed whole cell must be coated on solid phase. It is easy to coat lipopolysaccharide on ELISA plate but troublesome to purify it. As it is easy to obtain the killed whole cells, the development of the appropriate method by which those antigens of S. typhi are optimally coated on solid phase is needed. To establish the appropriate method, carbonate buffer, methanol or poly-L-lysine was applied as binding substance on polystyrene or polyvinylchloride plate as solid phase when the killed whole cell antigens of S. typhi varided as follows: $10^6$, $10^7$, $10^8$ and $10^9\;cell/ml$. The criteria of the optimal method were determined as follows: 1. The optical density of positive sera is above 1.0(0.6 in IgM) at 1:10 serum dilution and is 0.3(0.2 in IgM) higher than that of negative sera: 2. The O.D. of sera is flat or lowering according to serum dilution: 3. It must be that the O.D. of negative sera is lower than 0.2 at the point of serum dilution where the O.D. of positive sera is higher than 1.0(0.5 in IgM). The results obtained were summarized as follows: 1. The methods which fitted the above criteria were to use poly-L-lysine as binding substance, polyvinylchloride plate as solid phase and $10^7\;cell/ml$ as antigen concentration of S. typhi(poly-L-lysine/polyvinylchloride/$10^7$) and poly-L-lysine/polyvinylchloride/$10^8$ in detecting IgG antibody, methanol/polystyrene/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgM and carbonate buffer/polystyrene/$10^8$, carbonate buffer/polystyrene/$10^9$, methanol/polystyrene/$10^8$, methanol/polyvinylchloride/$10^8$, methanol/polyvinylchloride/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgA. 2. The coaling method using poly-L-lysine, polyvinylchloride plate and $10^8\;cell/ml$ was best to assay IgG, IgM and IgA antibody all in one. By this method, to assay the each immunoglobulin calss with an appropriate fixed serum dilution, 1:320 dilution was best.

• 생명과학회지
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• 제18권11호
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• pp.1507-1512
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• 2008

오랜 기간 동안의 연구에도 불구하고 사람에 특이적으로 장티푸스를 유발하는 S. typhi는 실험동물을 대상으로 하는 감염모델이 확립되어 있지 않기 때문에 S. typhi의 병원성 유발기작에 관한 정보는 부족하다. S. typhi Ty2 균주는 최소배지에서 시스테인의 영양요구성을 지닌다. 본 연구에서는 시스테인 영양요구성이, S. typhi Ty2 균주가 실험동물에서 균체형성을 하는데 어떤 영향을 미치는지를 조사하기 위해 시스테인 영양요구성을 상보할 수 있는 유전자를 찾고자 하였다. S. typhimurium의 genomic library로 형질전환된 S. typhi 균들 중 시스테인을 함유하지 않은 최소배지에서 생육을 하는 3개의 형질전환 균주를 선별하였으며, 이들 중 2개는 S. typhi의 시스테인 영양요구성을 아주 약하게 상보하였고 하나는 명확하게 시스테인의 영양요구성을 상보하였다. 이 클론에 포함되어져 있는 3개의 ORF의 시스테인 영양요구성을 분석한 결과, STM1490을 가진 클론이 S. typhi의 시스테인 영양요구성을 상보하였다. 비록 S. typhi에도 STM1490에 해당하는 유전자가 존재하지만 S. typhimurium의 STM1490에 해당하는 ORF와 비교하였을 때 2개의 아미노산 잔기가 서로 달랐다. 이들의 차이가 시스테인 영양요구성을 보이는 것은 아닌지 확인하기 위해 Overlapping PCR을 통해 S. typhimurium의 STM1490 아미노산(H229Y, C246W)을 치환하였다. 아미노산을 바꾼 돌연변이체도 역시 시스테인 영양요구성을 상보할 수 있어서 아미노산의 차이는 아닌 것을 확인되었으므로 그 외의 다른 인자들이 영양요구성에 관여하는 것으로 추정되었고 후속연구에서 그인자를 찾을 수 있을 것으로 생각된다.

• Biomolecules & Therapeutics
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• 제7권3호
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• pp.300-306
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• 1999

To study the antigenicity of BR92021(Vi polysaccharide typhoid vaccine), active systemic ana-phylaxis and passive cutaneous anaphylaxis were tested in guinea pigs. The groups were as follows: group I(low dose, 30 $\mu\textrm{l}$/kg), group II(high dose, 300 $\mu\textrm{l}$/kg), group III(300 $\mu\textrm{l}$/kg plus complete Freund's adjuvant), group IV(positive control, ovalbumin plus complete Freund's adjuvant) and group V(saline-treated control). Male Hartley guinea pigs at 7 weeks of age were sensitized subcutaneously with the test article or saline three times per week for three weeks(j.e., total 9 times). For groups III and IV, animals were sensitized subcutaneously with either the test article or ovalbumin plus complete Freund's adjuvant once per three week for 6 weeks(i.e., total 3 times). Twelve days after the last sensitization, the blood was collected from the sensitized animals for the passive cutaneous anaphylaxis test. In addition, the sensitized animals were subjected to the active systemic anaphylaxis test on fourteen days after the last sensitization by an intravenous challenge with either the test article or ovalbumin. In group I, mild(1/5) or moderate(4/5) symptoms of anaphylactic shock were observed. In group II, no sign(1/5), moderate(3/5) and severe(1/5) symptoms were observed. In group III, four animals of revealed moderate signs and one of 5 showed no signs of anaphylactic shock. In group IV, all 5 animals showed severe signs of shock. In group V, one of 5 revealed moderate and four of 5 showed no signs. The necropsy findings related to the active systemic anaphylaxis were observed in most animals of groups I to V In the passive cutaneous anaphylaxis test, the antiserum was diluted 10- to 5120- fold and was injected intradermally on the clipped back of recipient animals, followed by an intravenous challenge with either the test article or ovalbumin. No animals in groups I, II, III and V showed the positive reaction, whereas all animals in group IV, the positive control, showed the positive reaction at the dilution range of x1280 to x5120. Our results indicate that the test article, BR92021, may have weak antigenic potential in male guinea pigs.

• 대한미생물학회지
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• 제18권1호
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• pp.73-85
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• 1983

The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

• 대한미생물학회지
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• 제21권4호
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• pp.447-453
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• 1986

To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

• 한국식품위생안전성학회지
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• 제18권3호
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• pp.139-145
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• 2003

본 조사는 한국 소비자들의 식중독 경험여부, 식중독 원인균에 대한 지식 및 식품안전과 식중독 예방을 위한 행동에 있어서의 인식 정도를 알아보기 위하여 2001년 11월 일반성인 총 1,040명을 대상으로 전국적으로 전화조사를 실시하였다. 그 결과 12.4%는 연 1회 이상 식중독을 경험하고 있으며, 0.3%는 식중독으로 인해 병원에 입원하는 것으로 나타났다. 식중독이 발생한 원인장소로는 일반음식점에서의 음식 섭취가 37.2%로 가장 높게 나타났고, 가정에서의 부주의가 21.2% 등으로 나타났으며, 원인식품으로는 고기 등의 육류 및 가공품이 41.7%, 생선 등 어패류 및 가공품이 18.7% 등으로 나타났다. 식중독 원인균에 대한 인식에서는 콜레라(75.5%), 비브리오(73%), 이질(65.5%), 장티푸스(51.8%), 살모넬라증(47.5%)의 순으로 잘 알고 있는 것으로 나타났지만 리스테리아증(9.9%), 브루셀라중(8.3%)에 대해서는 대부분이 모르거나 음식물로 인해 발생하지 않는 것으로 인식하고 있는 것으로 나타났다. 한편, 조사자료를 바탕으로 3가지 모델로 구분 즉, Model 1은 식중독 원인균에 대한 지식, Model 2는 식품안전에 대한 인식, Model 3은 식중독 예방을 위한 행동에 대한 인식으로 구분하였고, 각 모델들은 조사자의 일반사항에서 나타난 변수와 식중독 경험여부 별로 다중회귀분석(Multiple regression analysis)을 수행하였다. 그 결과 식중독 원인균에 대한 인식(Model 1)은 여러 변수 중 교육수준(OR 0.536), 결혼상태(OR 0.529)가 영향요인으로 나타났으며, 특히, 과거 식중독 경험이 가장 중요한 영향요인으로 나타났다(OR 1.714). 식품안전에 대한 인식(Model 2)에서는 교육수준이 중요한 영향요인으로 나타났으며(OR 0.702), 식중독 예방을 위한 행동에 대한 인식(Model 3)에서는 교육수준(OR 0.816)과 성별(OR 0.650)이 중요한 영향으로 나타났으며, 과거 식중독 경험이나 식품안전에 대한 인식정도는 식중독 예방을 위한 행동에 대한 인식과는 상관관계가 없는 것으로 나타났다. 결론적으로 식품안전에 있어 경험과 지식은 실제 식중독 예방을 위한 행동에는 크게 영향을 미치지 않음을 의미하는 것으로 볼 수 있다.