• 식품과학과 산업
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• 제52권3호
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• pp.220-228
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• 2019

Rapid development of next-generation sequencing (NGS) technology is available to study microbes in genomic level. This NGS has been widely used in DNA/RNA sequencing for genome sequencing, metagenomics, and transcriptomics. The food microbiology area could be categorized into three groups. Food microbes including probiotics and food-borne pathogens are studied in genomic level using NGS for microbial genomics. While food fermentation or food spoilage are more complicated, their genomic study needs to be done with metagenomics using NGS for compositional analysis. Furthermore, because microbial response in food environments are also important to understand their roles in food fermentation or spoilage, pattern analysis of RNA expression in the specific food microbe is conducted using RNA-Seq. These microbial genomics, metagenomics, and transcriptomics for food fermentation and spoilage would extend our knowledge on effective utilization of fermenting bacteria for health promotion as well as efficient control of food-borne pathogens for food safety.

• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.83-95
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• 2023

These days, bacterial detection methods have some limitations in sensitivity, specificity, and multiple detection. To overcome these, novel detection and identification method is necessary to be developed. Recently, NGS panel method has been suggested to screen, detect, and even identify specific foodborne pathogens in one reaction. In this study, new NGS panel primer sets were developed to target 13 specific virulence factor genes from five types of pathogenic Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium, respectively. Evaluation of the primer sets using singleplex PCR, crosscheck PCR and multiplex PCR revealed high specificity and selectivity without interference of primers or genomic DNAs. Subsequent NGS panel analysis with six artificially contaminated food samples using those primer sets showed that all target genes were multi-detected in one reaction at 10⁸-10⁵ CFU of target strains. However, a few false-positive results were shown at 10⁶-10⁵ CFU. To validate this NGS panel analysis, three sets of qPCR analyses were independently performed with the same contaminated food samples, showing the similar specificity and selectivity for detection and identification. While this NGS panel still has some issues for detection and identification of specific foodborne pathogens, it has much more advantages, especially multiple detection and identification in one reaction, and it could be improved by further optimized NGS panel primer sets and even by application of a new real-time NGS sequencing technology. Therefore, this study suggests the efficiency and usability of NGS panel for rapid determination of origin strain in various foodborne outbreaks in one reaction.

• 환경생물
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• 제36권2호
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• pp.165-173
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• 2018

저어새의 먹이생물을 파악하기 위해 2010년 6월부터 2014년 6월까지 인천 남동유수지에서 저어새의 토사물 시료를 채집하여 현미경 관찰 및 차세대염기서열(NGS) 기법으로 분석하였다. 저어새의 먹이생물은 어류, 갑각류, 다모류, 곤충류로 구성되어 있었으며, 주로 저어새는 어류와 갑각류를 섭이하는 것으로 나타났다. 최우점 먹이생물은 풀망둑(Acanthogobius hasta)이었으며, 이 외에도 길게(Macrophthalmus abbreviates), 징거미새우류(Macrobrachium sp.), 칠게(Macrophthalmus japonicus), 각시흰새우(Exopalaemon modestus), 참갯지렁이(Neanthes japonica)가 우점 먹이생물로 출현하였다. 이들 먹이생물은 번식지 인근지역인 송도갯벌과 시화호에서 흔히 발견되며, 저어새는 채식지로써 이들 지역에 대한 의존도가 높을 것으로 판단된다. 현미경과 NGS로 분석한 일부 먹이생물에서 차이를 보였는데, 이는 토사물 내 먹이생물은 저어새의 위 내에서 분해되어 현미경 분석을 통한 형태학적 분류 특징을 찾기 어려웠던 반면, NGS 분석은 유전자를 통해 분류가 가능하기 때문에 형태학적 분석의 결과보다 높은 종 다양성을 보인 결과이다.

• 환경생물
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• 제36권2호
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• pp.131-139
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• 2018

기후 변화로 인한 수온의 상승은 어류 서식지에 영향을 미친다. 수온의 변화는 어류 생리 거의 모든 부분에 영향을 미치는 것으로 알려져 있다. 기후 변화에 따른 수온의 상승은 산소 용해도의 감소 및 산소 운반 헤모글로빈의 결합 능력의 감소로 인해 저산소증을 초래할 수 있다. 본 연구는 대서양 연어(Salmo salar) 치어 성장의 최적수온($15^{\circ}C$)보다 고수온($20^{\circ}C$)에 사육 시, 대서양 연어 치어의 건강상태를 평가하기 위해 수행되었다. 평가 방법은 NGS RNAseq 분석방법을 이용하여 생체지표유전자를 개발하고, RT-qPCR 분석을 이용하여 생체지표유전자의 발현양상을 조사하는 것이다. 개발한 생체지표유전자로는 interferon alpha-inducible protein 27-like protein 2A transcript variant X3, protein L-Myc-1b-like, placenta growth factor-like transcript variant X1, fibroblast growth factor receptor-like 1 transcript variant X1, transferrin, intelectin, thioredoxin-like, c-type lectin lectoxin-Thr1-like, ladderlectin-like 및 calponin-1 등이다. 선택된 생체지표 유전자는 NGS RNAseq 분석을 통해 수온변화에 민감하게 발현한 유전자들이며, RT-qPCR 분석을 통한 이들 유전자의 발현 양상은 NGS RNAseq 분석을 통한 발현 양상과 매우 유사하게 나타났다.

• 한국축산시설환경학회지
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• 제21권3호
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• pp.93-98
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• 2015

The objective of this study was to review application of next-generation sequencing (NGS) to investigate microbiome in the livestock sector. Since the 16S rRNA gene is used as a phylogenetic marker, unculturable members of microbiome in nature or managed environments have been investigated using the NGS technique based on 16S rRNA genes. However, few NGS studies have been conducted to investigate microbiome in the livestock sector. The 16S rRNA gene sequences obtained from NGS are classified to microbial taxa against the 16S rRNA gene reference database such as RDP, Greengenes and Silva databases. The sequences also are clustered into species-level OTUs at 97% sequence similarity. Microbiome similarity among treatment groups is visualized using principal coordinates analysis, while microbiome shared among treatment groups is visualized using a venn diagram. The use of the NGS technique will contribute to elucidating roles of microbiome in the livestock sector.

• 대한의생명과학회지
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• 제22권4호
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• pp.150-159
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• 2016

Knowledge of the distribution and biodiversity of environmental bacteria and the ecosystem that influences them is crucial for predicting an ecosystem. However, bacterial culture methods can only analyze approximately 0.1% of the existing microorganisms, those that are readily cultured under laboratory conditions. By contrast, next-generation sequencing (NGS) has generally been known to obtain more diverse profiling of bacterial composition. We compared the bacterial communities using both a culture-dependent (MALDI-TOF) and culture-independent (NGS) methods. Environmental specimens were obtained from both freshwater and seawater. Water samples were also analyzed by both pyrosequencing and MiSeq sequencing, in order to select one NGS platform which could analyze comparatively more diverse microbiota. Bacterial distribution analyzed with MALDI-TOF showed no difference between the microbiota of freshwater and seawater, whereas the results analyzed with NGS distinguished between the two. The diversity indexes of MiSeq sequencing were higher than for Pyrosequencing. This indicated that MiSeq sequencing is capable of analyzing a comparatively wider diversity of bacteria. The genus of Flavobacterium and Planktophila were identified as being unique to freshwater, whereas EU801223 and OM43 were found in the seawater. Difference between the bacterial composition of the freshwater and seawater environments was identified by MiSeq sequencing analysis.

• Genomics & Informatics
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• 제11권4호
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• pp.191-199
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• 2013

High-throughput next-generation sequencing (NGS) technology produces a tremendous amount of raw sequence data. The challenges for researchers are to process the raw data, to map the sequences to genome, to discover variants that are different from the reference genome, and to prioritize/rank the variants for the question of interest. The recent development of many computational algorithms and programs has vastly improved the ability to translate sequence data into valuable information for disease gene identification. However, the NGS data analysis is complex and could be overwhelming for researchers who are not familiar with the process. Here, we outline the analysis pipeline and describe some of the most commonly used principles and tools for analyzing NGS data for disease gene identification.

• 한국동물위생학회지
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• 제39권1호
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• pp.41-49
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• 2016

In the present study, fast and robust methods for the next generation sequencing (NGS) were developed for analysis of PRRSV full genome sequences, which is a positive sensed RNA virus with a high degree of genetic variability among isolates. Two strains of PRRSVs (VR2332 and VR2332-R) which have been maintained in our laboratory were used to validate our methods and to compare with the sequence registered in GenBank (GenBank accession no. EF536003). The results suggested that both of strains had 100% coverage with the reference; the VR2332 had the coverage depth from minimum 3 to maximum 23,012, for the VR2332-R from minimum 3 to maximum 41,348, and 22,712 as an average depth. Genomic data produced from the massive sequencing capacities of the NGS have enabled the study of PRRSV at an unprecedented rate and details. Unlike conventional sequence methods which require the knowledge of conserved regions, the NGS allows de novo assembly of the full viral genomes. Therefore, our results suggested that these methods using the NGS massively facilitate the generation of more full genome PRRSV sequences locally as well as nationally in regard of saving time and cost.

• 한국항행학회논문지
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• 제13권6호
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• pp.957-963
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• 2009

본 논문은 스토리지 디바이스에 대한 자원관리 시스템의 설계에 관한 것이다. 컴퓨팅 환경의 변화와 복잡화에 따라 자원에 대한 효율적인 관리방안으로 제안되고 있는 표준관리응용프로그램을 설계하기 위해서 객체지향 모델링 기술에 바탕을 둔 공통정보모델을 사용하여 관리객체에 대한 모델링 과정을 수행하였다. 표준화된 관리응용프로그램 구현 방법을 사용하여 사용자에게 시각적인 특성을 제공할 수 있도록 GUI인터페이스를 구현하였으며 시스템의 구성 요소를 자동으로 검색, 관리하고 각 구성 요소의 상태 정보를 주의/경고/긴급의 3단계로 분류하여 사용자에게 제공하였다. 또한 다양한 구성 요소의 운용을 위한 설정기능을 제공하는 관리응용시스템을 구현하였다.

• 식품과학과 산업
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• 제50권1호
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• pp.2-10
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• 2017

Human have eaten various traditional fermented foods for a numbers of million years for health benefit as well as survival. The beneficial effects of fermented foods have been resulted from complex microbial communications within the fermented foods. Therefore, the holistic approaches for individual identification and complete microbial profiling involved in their communications have been of interest to food microbiology fields. Microbiome is the ecological community of microorganisms that literally share our environments including foods as well as human body. However, due to the limitation of culture-dependent methods such as simple isolations of just culturable microorganisms, the culture-independent methods have been consistently developed, resulting in new light on the diverse non-culturable and hitherto unknown microorganisms, and even microbial communities in the fermented foods. For the culture-independent approaches, the food microbiome has been deciphered by employing various molecular analysis tools such as fluorescence in situ hybridization, quantitative PCR, and denaturing gradient gel-electrophoresis. More recently, next-generation-sequencing (NGS) platform-based microbiome analysis has been of interest, because NGS is a powerful analytical tool capable of resolving the microbiome in respect to community structures, dynamics, and activities. In this overview, the development status of analysis tools for the fermented food microbiome is covered and research trend for NGS-based food microbiome analysis is also discussed.