• Genomics & Informatics
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• 제19권3호
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• pp.34.1-34.6
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• 2021

Digital PCR (dPCR) is the third-generation PCR that enables real-time absolute quantification without reference materials. Recently, global diagnosis companies have developed new dPCR equipment. In line with the development, the Lab On An Array (LOAA) dPCR analyzer (Optolane) was launched last year. The LOAA dPCR is a semiconductor chip-based separation PCR type equipment. The LOAA dPCR includes Micro Electro Mechanical System that can be injected by partitioning the target gene into 56 to 20,000 wells. The amount of target gene per wells is digitized to 0 or 1 as the number of well gradually increases to 20,000 wells because its principle follows Poisson distribution, which allows the LOAA dPCR to perform precise absolute quantification. LOAA determined region of interest first prior to dPCR operation. To exclude invalid wells for the quantification, the LOAA dPCR has applied various filtering methods using brightness, slope, baseline, and noise filters. As the coronavirus disease 2019 has now spread around the world, needs for diagnostic equipment of point of care testing (POCT) are increasing. The LOAA dPCR is expected to be suitable for POCT diagnosis due to its compact size and high accuracy. Here, we describe the quantitative principle of the LOAA dPCR and suggest that it can be applied to various fields.

• 환경생물
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• 제41권4호
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• pp.657-665
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• 2023

The root-lesion nematode Pratylenchus spp. is the most important plantparasitic nematode due to its worldwide distribution, wide host ranges, and migratory endoparasitic characteristics. One population of Pratylenchus collected from the giant pussy willow (Salix chaenomeloides Kimura) in the Andong area as part of a nematode survey in Korea was characterized morphologically and by molecular methods. The analysis of morphological measurements and morphometric characteristics, as well as DNA sequencing of the rRNA large subunit (LSU) D2/D3 expansion segments and the internal transcribed spacer (ITS) gene sequence, confirmed the identity of this population as P. hippeastri. This study is the first report of P. hippeastri associated with Salix chaenomeloides in Korea and worldwide. Further studies on distribution and pathogenicity in different P. hippeastri host crops, such as grapevines, strawberries, and apples, are necessary. The taxonomic keys to 16 Pratylenchus species in Korea are provided.

• Clinical and Experimental Pediatrics
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• 제48권8호
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• pp.871-876
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• 2005

목 적 : 한국 소아 1형 당뇨병에서 TNF promoter -857T/C와 -1031C/T 및 $LT-{\alpha}$ 유전자 다형성과 질병감수성과의 관련성을 평가하고자 하였다. 방 법 : 1형 당뇨병으로 진단 받은 소아 49명(여아 29명, 남아 20명)과 정상 대조군 94명의 혈액을 채취하여 DNA를 추출하였다. 추출한 DNA에 대하여 allele-specific PCR법을 이용하여 TNF promotor -1031C/T 다형성을, PCR-RFLP법을 이용하여 TNF promotor -857T/C, $LT-{\alpha}$ 유전자 다형성을 분석하였다. 결 과 : 환자군과 대조군 사이에서 TNF promoter -857T/C, -1031C/T 다형성의 분포는 차이가 없었다. 환자들의 임상적 특징에 따라 아군(subgroup)으로 분류하였을 때, 진단 시 당뇨병성 케톤산혈증으로 발현한 환자들에서 TNF promoter -1031C/T 다형성의 TT 유전자형의 빈도가 당뇨병성 케톤산혈증으로 발현하지 않은 환자들과 비교하여 유의하게 낮았다(P<0.05). 다른 임상적 특성들과 이들 유전자 다형성 사이에는 관련성이 없었다. 또 환자와 대조군 사이에 $LT-{\alpha}$ 유전자 다형성의 분포는 차이가 없었으며, 임상적 특성과의 관련성도 없었다. 결 론 : 이 연구를 통하여 TNF promoter -857T/C, $LT-{\alpha}$ 유전자 다형성이 한국 소아에서 1형 당뇨병의 질병감수성과 관련이 없음을 알 수 있었다. 그러나 TNF promoter -1031C/T 다형성은 당뇨병성 케톤산혈증과 같은 1형 당뇨병의 특정 임상양상에 영향을 미칠 수 있는 유전적 인자로 생각된다.

• Parasites, Hosts and Diseases
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• 제52권3호
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• pp.325-329
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• 2014

The phylogenetic relationships of the 3 Neodiplostomum spp. (Digenea: Neodiplostomidae) occurring in Korea (N. seoulense, N. leei, and N. boryongense) were analyzed using the partial mitochondrial cytochrome c oxidase subunit 1 (CO1) gene. The adult flukes were recovered from Sprague-Dawley rats (N. seoulense) and newborn chicks (N. leei and N. boryongense) experimentally infected with the neodiplostomula from the grass snake, Rhabdophis tigrinus tigrinus. The genomic DNA was amplified using specific primers, and the sequence of CO1 was obtained. According to the results, the pairwise similarity was 96.1% between N. boryongense and N. seoulense, but was 95.0% between N. boryongense and N. leei and 94.2% between N. leei and N. seoulense. The results demonstrated a closer phylogenetic relationship between N. seoulense and N. boryongense. This high relationship of N. seoulense and N. boryongense may be related to their similar morphologic features including the limited distribution of vitellaria and the presence of a genital cone. N. leei is distinct on the other hand with an extensive distribution of vitellaria and the absence of a genital cone.

• 대한수의학회지
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• 제39권1호
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• pp.27-33
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• 1999

In the mammalian ovary, follicular development and atresia continuously occur during the reproductive cycles. Follicular atresia occurs through granulosa cell apoptosis. Apoptosis is known as the physiological cell death, which is regulated by bcl-2 gene family. In the bcl-2 gene family, bcl-2 and bcl-xLong are known as inhibitors of apoptosis, whereas bax and bcl-xShort are known as inducer of apoptosis. We thought that bcl-2 protein is associated with follicular development and atresia. But it is not known that the distribution of cells containing bcl-2 protein during follicular development and atresia. Therefore, to examine the distribution of cells with bcl-2 protein during ovarian follicular development and atresia, the immunohistochemistry was used in the rat ovary. Bcl-2 immunoreactivity was localized in the interstitial cells, theca externa cells and granulosa cells around of antrum. All positive signals were observed in the cytoplasm of these cells. Positive signals were strongly observed in the interstitial and theca externa cells of growing antral follicles. While, positive signals were weakly observed in these cells from atretic antral follicles. Positive signals were very weakly observed in the granulosa cells of growing and atretic antral follicles. According to these data, we suggested that bcl-2 proteins which were strongly expressed in the interstitial cells and theca externa cells of growing antral follicles inhibit follicular atresia. And we purposed that bcl-2 proteins regulated follicular development and atresia through the action of bcl-2 gene family.

• Genomics & Informatics
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• 제8권2호
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• pp.76-80
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• 2010

Although autism spectrum disorder (ASD) has been thought to have a substantial genetic background, major contributing genes have yet to be identified or successfully replicated. Immunological dysfunction has been suggested to be associated with ASD, and T cell-mediated immunity was considered important for the development of ASD. In this study, we analyzed 163 ASD subjects and 97 normal controls by genomic quantitative PCR to evaluate the association between the copy number variation of the 7q34 locus, harboring the TCRB gene, and ASDs. As a result, there was no significant difference of the frequency distribution of TCRB copy numbers between ASD cases and normal controls. TCRB gene copy numbers ranged from 0 to 5 copies, and the frequency distribution of each copy number was similar between the two groups. The proportion of the individuals with <2 copies of TCRB was 52.8% (86/163) in ASD cases and 57.1% (52/91) in the control group (p=0.44). The proportion of individuals with >2 copies of TCRB was 11.7% (19/163) in ASD cases and 12.1% (11/91) in the control group (p=0.68). After the effects of sex were adjusted by logistic regression, ORs for individuals with <2 copies or >2 copies showed no significant difference compared with the diploid copy number as reference (n=2). Although we could not see the positive association, our results will be valuable information for mining ASD-associated genes and for exploring the role of T cell immunity further in the pathogenesis of ASD.

• 한국미생물·생명공학회지
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• 제47권2호
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• pp.310-316
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• 2019

Distribution of Salmonella enterica serovars and their associated virulence determinants is wide-spread among food animals, which are continuously implicated in periodic salmonellosis outbreaks globally. The aim of this study was to determine and evaluate the diversity of five Salmonella serovar virulence genes (invA, pefA, cdtB, spvC and iroN) isolated from food animals and humans. Using standard microbiological techniques, Salmonella spp. were isolated from the feces of humans and three major food animals. Virulence determinants of the isolates were assayed using PCR. Clonal relatedness of the dominant serovar was determined via pulsed-field gel electrophoresis (PFGE) using the restriction enzyme, Xbal. Seventy one Salmonella spp. were isolated and serotyped into 44 serovars. Non-typhoidal Salmonella (NTS; 68) accounted for majority (95.8%) of the Salmonella serovars. Isolates from chicken (34) accounted for 47.9% of all isolates, out of which S. Budapest (14) was predominant (34.8%). However, the dominant S. Budapest serovars showed no genetic relatedness. The invA gene located on SPI-1 was detected in all isolates. Furthermore, 94% of the isolates from sheep harbored the spvC genes. The iroN gene was present in 50%, 100%, 88%, and 91% of isolates from human, chicken, sheep, and cattle, respectively. The pefA gene was detected in 18 isolates from chicken and a single isolate from sheep. Notably, having diverse Salmonella serovars containing plasmid encoded virulence genes circulating the food chain is of public health significance; hence, surveillance is required.

• 한국어병학회지
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• 제35권1호
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• pp.47-56
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• 2022

Lincomycin as one of the lincosamides antibiotics have been mainly used in human and livestock fields, but have not been used in aquaculture. In this study, the distribution of minimum inhibitory concentration (MIC) values against lincomycin and the detection of the macrolide-lincosamide-streptogramin (MLS) resistance gene were confirmed in bacterial pathogens isolated from cultured olive flounder (Paralichthys olivaceus). Of the 107 strains isolated from Jeju, 36 strains of Gram-positive bacteria and 71 strains of Gram-negative bacteria were identified. Most of Streptococcus spp. was found to have a MIC value of less than or equal to 0.5 ㎍/mL, and Edwardsiella piscicida was found to have a MIC value higher than 1,024 ㎍/mL. V. harveyi and V. alginolyticus mostly showed MIC values of 256 ㎍/mL, but V. scophthalmi displayed values of 8~64 ㎍/mL. In the detection of MLS resistance gene, erm(B) was detected in 9 strains of Streptococcus spp., and erm(A) was confirmed in one strain.

• 한국발생생물학회지:발생과생식
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• 제27권2호
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• pp.91-99
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• 2023

The sea cucumber, Apostichopus japonicus, is one of the most valuable aquatic species. The color of body wall and appearance are important for the value of sea cucumbers. To examine expression pattern of long-chain acyl-coenzyme A dehydrogenase (LCAD), nuclear distribution C-containing protein 3 (NUDCD3), and receptor tyrosine kinase Tie-1 (TIE1), previously reported as differently expressed genes during the pigmentation of sea cucumber, we analyzed the temporal profiles of LCAD, NUDCD3, and TIE1 mRNAs in LED-exposed and light-shielded A. japonicus. Real-time quantitative PCR revealed that the LCAD, NUDCD3, and TIE1 mRNAs from the juveniles at 40-60 days post-fertilization (dpf) exhibited increasing patterns as compared to those of an early developmental larva (6-dpf). At 60-dpf juveniles, the LCAD and TIE1 mRNA levels of LED-exposed individuals were higher than those of light-shielded ones, whereas at 40-dpf and 50-dpf juveniles, the NUDCD3 mRNA expression was higher in the light-shielded condition (p<0.05). In the pigmented juveniles (90-dpf), the LCAD and TIE1 mRNA levels tended to show higher levels in red individuals than those in green ones, but there was a conversely higher level of NUDCD3 mRNA in green larva. In situ examination of LCAD and NUDCD3 mRNAs in light-shielded 6-dpf larva revealed that both genes are mainly expressed in the internal organs compared to the body surface. Together, these results may provide insights into the differential gene expression of LCAD, NUDCD3, and TIE1 during pigmentation process of the sea cucumber.

• 농업과학연구
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• 제22권2호
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• pp.170-179
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• 1995

본 연구는 축협중앙회 한우개량사업소에서 사육하고 있는 한우 238두의 혈청내에 존재하는 혈액 단백질 및 효소의 유전적 다형을 조사하기 위하여, 혈청단백질인 post-transferrin-2(pTf-2), transferrin(Tf), post-albumin(pAlb) 및 albumin(Alb)과 혈청 효소인 ceruloplasmin(Cp) 및 amylase-I(Am-I), 그리고 혈구단백질인 hemoglobin(Hb)을 PAGE(polyacrylamide gel electrophoresis)와 STAGE(starch gel electrophoresis) 방법으로 유전적 다형을 분석하여 이들의 유전자형 및 유전자빈도를 추정하였던바 그 결과는 다음과 같다. 1. 혈청단백질인 pTf-2 좌위는 pTf-2 F와 S의 공우성 대립유전자가 검출되었고, 유전자형은 pTf-2 FF, FS 및 SS 형이 확인되었으며, 이들의 분포는 각각 46.22, 46.64 및 7.14%이었고, 유전자 빈도는 pTf-2 F와 S에서 각각 0.695 및 0.305이었다. 2. Tf 좌위는 Tf A, D1, D2 및 E의 대립유전자가 검출되었으며, 유전자형에 있어서는 Tf AA, ADl, AD2, AE, D1D1, D1D2, D2D2 DIE D2E 및 EE형이 확인되었고, 이들의 분포는 각각 0.84, 13.87, 13.03, 10.92, 22.27, 12.61, 2.94, 15.l3, 6.72 및 1.68%이었으며, 유전자빈도에 있어서는 Tf A, D1, D2 및 E의 빈도가 각각 0.197, 0.430, 0.191 및 0.181이었다. 3. pAlb 좌위는 pAlb F와 S 유전자가 검출되었으며, 유전자형의 분포에 있어서는 pAlb FFFS 및 SS 형이 각각 42.86, 33.19 및 23.95%이었고, 유전자빈도에 있어서는 pAlb F와 S에서 각각 0.595 및 0.405이었으며, Alb에서는 Alb A 유전자 빈도가 1.000이었다. 4. Cp 좌위는 Cp F와 S 유전자가 확인되었으며, 유전자형의 분포는 Cp FF, FS 및 SS 형에서 각각 23.11, 34.87 및 42.02% 이었고, Cp F와 S 유전자빈도는 각각 0.405 및 0.595이었다. 5. Am-I 좌위는 Am-I B와 C 대립유전자가 검출되었으며, 유전자형의 분포는 Am-I BB, BC 및 CC형에서 각각 51.26, 16.81 및 31.93%이었고, 유전자빈도는 Am-I B와 C가 각각 0.597 및 0.403이었다. 6. Hb 좌위는 Hb A와 B 대립유전자가 검출되었으며, 유전자형의 분포는 Hb AA, AB 및 BB형에서 각각 83.19, 16.39 및 0.42%이었고, Hb A와 B의 유전자빈도는 각각 0.914 및 0.086%이었다.