• Title/Summary/Keyword: DNA-index

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The Assessment of Ultrasensitive HBsAg kit's Sensitivity level and Performance in Detection of Mutant Forms (Ultra-sensitive HBsAg IRMA 키트의 민감도 및 변이형 검출능 평가)

  • Ha, Dong-Hyuk;Min, Kyung-Sun;Noh, Gyeong-Woon;Kim, Hyun-Ju
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.121-125
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    • 2011
  • Purpose: The DNA-type virus HBV, discovered by D. Dane and others in 1976, is approximately 42nm big and known as the main cause of liver-related diseases around the world. HBsAg has 4 kinds of subtypes including adw, adr, ayw and ayr and besides common antigen factor a, there are d, y, r, w. From the methods of serologically testing HBV, IRMA, EIA and CLIa were developed for testing HBsAg and are being used in examining the surface antigen of HBV. In this study, among the methods for testing HBV, the recently developed RIAKEY Ultrasensitive HBsAg IRMA kit's sensitivity level and performance in detection of mutant forms were measured and compared with CLIA. Materials and methods: Two certified reference materials, which are WHO 1st International Standard 1985(80/549) and WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA), were used in the examination and the sensitivity level was measured by diluting these materials from 0.08 IU/ml to 0.005 IU/ml. The materials for examining the detection of mutant forms included 9 kinds of subtype 'ad' and one kind of subtype 'ay' purchased from DSI company. Also, with the use of positive and negative samples, they was compared with CLIA. Result: Ultrasensitive HBsAg kit based on IRMA method showed the detection of up to 0.01 IU/ml not only for WHO 1st International Standard 1985(80/549) but also for WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA) and the sensitivity level was measured as 0.01 IU/ml by WHO standard. In testing the performance for detection of mutant forms, the 9 kinds of subtype 'ad' and one kind of subtype 'ay' mutant materials were detected, demonstrating the capacity of detecting various types of mutant forms. Conclusions: With the clinical importance of sensitivity level and performance in detection of mutant forms increasing in the field of HBsAg diagnosis, the examination of IRMA's effectiveness using RIA method in the aspects of the sensitivity level and performance in detection of mutant forms was carried out and its result is as follows. The sensitivity level was measured as 0.01 IU/ml by WHO standard and it was possible to measure various types of mutant forms with high sensitivity. Thus it is suggested that more speedy and accurate reports could be produced from a nuclear medicine laboratory for clinical practitioners requiring results of various situations.

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Analysis of Genetic Diversity of Apple Cultivars Using RAPD and SSR Markers (RAPD와 SSR 마커를 이용한 사과 품종의 유전적 다양성 분석)

  • Cho, Kang-Hee;Heo, Seong;Kim, Jeong-Hee;Shin, Il Sheob;Han, Sang Eun;Kim, Se Hee;Kim, Dae-Hyun;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.525-533
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    • 2010
  • In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity of 34 Korean bred and introduced apple cultivars. Thirty-seven RAPD primers detected a total of 193 polymorphic bands (36.2%) with an average of 5.6. Twenty-six SSR markers generated a total of 112 alleles with an average 4.3 alleles per locus. Genetic diversity of 34 cultivars estimated by polymorphic information content (PIC) value ranged from 0.536 (CH03d12) to 0.952 (CH04c06) with an average of 0.843. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 305 polymorphic bands, the apple cultivars were classified four groups by similarity index of 0.640. The 'Seokwang' was included in group I. Group II consisted of 12 cultivars which have 'Golden Delicious' in their pedigree, with the exception of 'Spur Earliblaze' and 'Jonathan'. Group III included 13 cultivars which have usually 'Fuji' in their ancestry and bud sport of 'Fuji' cultivars. Group IV consisted of 8 cultivars with 'Hongro', 'Gamhong', and 'Saenara'. Similarity values among the tested apple cultivars ranged from 0.529 to 0.987, and the average similarity value was 0.647. The similarity index was the highest (0.987) between 'Hwarang' and 'Danhong', and the lowest (0.529) between 'Seokwang' and 'Hwarang'. The genetic relationships among the 34 studied apple cultivars were basically consistent with the known pedigree.

An Epidemiological Investigation on an Outbreak of Shigellosis in a Special School for Handicapped in Yeongcheon-si and in a Rehabilitation Facility in Gyeongsan-si, Korea, 2008 (2008년 영천시 지적장애인 학교와 경산시 재활원에서 발생한 세균성이질에 관한 역학조사)

  • Lee, Hyun-Dong;Lee, Soon-Ok;Lim, Hyun-Sul
    • Journal of agricultural medicine and community health
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    • v.34 no.1
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    • pp.24-33
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    • 2009
  • Objectives: In September 2008, an outbreak of diarrhea occurred among students attending Y school in Yeongcheon-si. Shigella sonnei was cultured from some of the rectal swabs. An epidemiological investigation was carried out to determine the source of the infection and the mode of transmission of the shigellosis outbreak. Methods: The index case lived in the D rehabilitation facility in Gyeongsan-si and an additional epidemiological investigation was carried out there. The cases could not be questioned due to their mental handicaps. The teachers were interviewed instead. A patient case was defined as a resident with diarrhea more than one time a day from September 18 to September 26, 2008 or a resident with confirmed Shigella sonnei at the Y school or the D rehabilitation facility. Results: The attack rate was 1.2% (8 persons) among 659 persons in the Y school and D rehabilitation facility. Five persons were microbiologically confirmed to have the infection and three persons were diagnosed on the basis of symptoms. Shigella sonnei was cultured from five of the 659 rectal swabs. However, 80 environmental specimens including drinking water, preserved foods, and cooking utensils were negative. All eight patients were Y school students and had been living in group boarding and lodging. Six of them lived in the D rehabilitation facility and two lived in the dormitory at the Y school. Five cases showed pulsed-field gel electrophoresis patterns that were identical for Shigella sonnei. Conclusions: The results of this study showed that the infection source of the shigellosis outbreak, in the two places, were identical. It is likely that the infections initially spread from a teacher or volunteer and then among the students.

Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation

  • Jeong, Chang-Dae;Mamuad, Lovelia L.;Kim, Seon-Ho;Choi, Yeon Jae;Soriano, Alvin P.;Cho, Kwang Keun;Jeon, Che-Ok;Lee, Sung Sil;Lee, Sang-Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.50-57
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    • 2015
  • This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

Analysis of cytosine adenine repeat polymorphism of the IGF-I promoter gene in children with idiopathic short stature (특발성 저신장증 환자에서 IGF-I 프로모터 cytosine-adenine repeat 유전자 다형성의 분석)

  • Moon, Jae Hoon;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.52 no.3
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    • pp.356-363
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    • 2009
  • Purpose : A polymorphism in the IGF-I gene promoter region is known to be associated with serum IGF-I levels, birth weight, and body length, suggesting that IGF-I gene polymorphism might influence postnatal growth. The present study aimed to investigate the role of this polymorphic cytosine-adenine (CA) repeat of the IGF-I gene in children with idiopathic short stature. Methods : The study involved 131 children (72 boys and 59 girls) diagnosed with idiopathic short stature, aged 715 years. Genomic DNA was extracted from anticoagulated peripheral whole blood. The primers were designed to cover the promoter region containing the polymorphic CA repeat. Data were analyzed using GeneMapper software. The correlations between age and serum IGF-I levels were analyzed using Spearmans correlation coefficient. Results : The CA repeat sequences ranged from 15 to 22, with 19 CA repeats the most common with an allele frequency of 40.6%. Homozygous for 19 CA repeat was 13.0%, heterozygous for 19 CA repeat was 56.5%, and 19 CA non-carrier was 30.5%. The three different genotype groups showed no significant differences in height, body weight and body mass index, and serum IGF-I levels. The serum IGF-I level and age according to the IGF-I genotypes were significantly correlated in the entire group, 19 CA repeat carrier group, and the non-carrier group. The three groups also showed no significant differences in the first year responsiveness to GH treatment. Conclusion : There were no significant different correlations between 19 CA repeat polymorphism and serum IGF-I levels according to genotype. Our results suggest that the IGF-I 19 CA repeat gene polymorphism is not functional in children with idiopathic short stature.

Effects of γ-Irradiation from Cobalt-60 on Immunogenicity of Eimeria tenella (Cobalt-60 γ-선 조사가 Eimeria tenella 의 면역원성에 미치는 영향)

  • Youn, Hee-jeong;Kang, Yung-bai;Jang, Du-hwan
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.657-664
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    • 1993
  • To reveal the immunogenicity of ${\gamma}-irradiated$ E tenella and its progeny, a series of experiments on the effects of Cobalt-to ${\gamma}-irradiation$ was performed. The SPF chickens inoculated with diffenrt doses of inoculum were challenged with $1{\times}10^5$ oocysts of virulent E tenella. The levels of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and of inoculum with $1{\times}10^4$ oocysts were recognized as proper as immunogen by comparison of survival rates, body weight gains, blood in feces and lesion scores in the chickens. In these trials of challenge with virulent E tenella after inoculation with $1{\times}10^4$ oocysts of the ${\gamma}-irradiated$ E tenella and its progeny, the survival rates of the chickens challenged with the virulent E tenella after immunization with the 1st and the 3rd progeny groups of ${\gamma}-irradiated$ E tenella oocysts were higher(l00%) than that(87.0%) of the challenged control group. The signs of blood in feces and the lesion scores were seen markedly lower with the ourput of the smaller number of oocysts, i.e. OPG 103,900 and 25,800 in the groups of the 1st and the 3rd progeny, respectively, than those(OPG 1,658,900) of the challenged control group. The body weight gains of the 1st and the 3rd progeny groups, the 1st week and the 2nd week after challenge, were higher (2.6g and 155.4g, 11.6g and 168.9g respectively) than those(-85.8g and 63.6g, respectively) of the challenged control group, and the feed conversion ratios(FCR 3.28 and 2.96) of the 1st and the 3rd progeny groups were lower than that(FCR 5.60) of the groups challenged control group. The anticoccidial indices(70.5 and 93.9) of the groups challenged with the virulent oocysts of E tenella after immunization with the 1st and the 3rd progeny of the ${\gamma}-irradiated$ E tenella were significantly higher than that (ACI -81.9) of the challenged control group. It was thought that the immunogenicity of ${\gamma}-irradiated$ E tenella would be increase according to increase the number of generation passaged in chicken. That might be because of increasing the pathogenicity of ${\gamma}-irradiated$ E tenella according to increase the number of generation passaged in chicken.

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Development of Method using LC-ESI-MS/MS and KASP for Identification of Gymnema sylvestre in Food (식품에서 당살초 판별을 위한 LC-ESI-MS/MS 분석법과 KASP 마커 개발)

  • Park, Boreum;Lee, Sun Hee;Eom, Kwonyong;Noh, Eunyoung;Moon Han, Kyoung;Hwang, Jinwoo;Kim, Hyungil;Baek, Sun Young
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.46-54
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    • 2022
  • Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.