• Title/Summary/Keyword: Identification and Detection

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MLPA Applications in Genetic Testing (유전자진단에 있어서 Multiplex Ligation Dependent Probe Amplification (MLPA)의 이론과 실제)

  • Kim, Gu-Hwan;Lee, Beom-Hee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.6 no.2
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    • pp.146-154
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    • 2009
  • Multiplex ligation dependent probe amplification (MLPA) is a PCR-based method to detect gene dosage. Since its introduction, MLPA has been used to test a large number of genes for major deletions or duplications. Genetic testing, as a diagnostic tool for genetic disease, has been used primarily to identify point mutations, including base substitutions and small insertions/deletions, using PCR and sequence analysis. However, it is difficult to identify large deletions or duplications using routine PCR- gel based assays, especially in heterozygotes. The MLPA is a more feasible method for identification of gene dosage than another routine PCR-based methods, and better able to detect deleterious deletions or duplications. In addition to detection of gene dosage, MLPA can be applied to identify methylation patterns of target genes, aneuploidy during prenatal diagnoses, and large deletions or duplications that may be associated with various cancers. The MLPA method offers numerous advantages, as it requires only a small amount of template DNA, is applicable to a wide variety of applications, and is high-throughput. On the other hand, this method suffers from disadvantages including the possibility of false positive results affected by template DNA quality, difficulties identifying SNPs located in probe sequences, and analytical complications in quantitative aspects.

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Comparison of Enzyme-linked Immunosorbent Assay with Reverse Transcription-polymerase Chain Reaction for Identification of Rotavirus in Neonates (신생아 로타바이러스 감염 진단에서 효소면역법과 중합효소연쇄반응과의 비교)

  • Kim, Sung Eun;Kim, Mi Ok;Park, Sun Young;Jung, Won Jo;Ma, Sang Hyeok;Kim, Yun Jung;Kim, Sun Ju
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.113-119
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    • 2000
  • Purpose : Rotavirus is a most common etiologic agent of pediatric gastroenteritis. The standard method to diagnose rotavirus infection was the detection of viral particles in specimens through electron microscopy. But it was complex. Enzyme immunoassay and latex agglutinin are preferred because they are relatively handy, inexpensive and take a short time, in comparison with electron microscopy. However, several reports have shown that the use of ELISA to diagnose rotavirus infection in neonates can result in false positive reactions. The main purpose of this study is to compare ELISA and RT-PCR in the diagnosis of neonatal rotavirus infection. Methods : Data presented in this study were obtained form 123 newborn babies in the nursery of the Fatima Hospital, Masan, Korea, form Jury to December, 1997. We obtained two samples of stool from each of the newborn babies and then performed the Rotazyme test and the RT-PCR. In the Rotazyme test, the results were interpreted according to visual findings. The samples were used for the RT-PCR test after at stock $-30^{\circ}C$ to identify rotavirus group A. The result of the two tests were compared. Results : The informations are divided into 73 males and females. Out of the total informations 15 were transferred from other hospitals. Their average gestational age was $38.5{\pm}1.6$ weeks. The average birth weight was $3134.8{\pm}539gm$. In the Rotazyme test, 75 samples turned out to be positive. Out of them, 55 samples(75.3%) were positive and 18 samples(24.7%) were negative in the RT-PCR. On the other hand, in the Rotazyme test, 50 samples turned out be negative. Out of them, 27 samples(54%) were positive and 23 samples(46%) were negative in the RT-PCR. Conclusion : Rotavirus infection in uncommon in neonates. The diagnosis based on visual findings using Rotazyme test has a disadvantage in the sense that it can result in false positive reactions and false negative reactions in the diagnosis of neonatal rotavirus infection.

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Identification of Hanwoo (Korean Native Cattle) Beef in Restaurants using Real-time PCR (시중 음식점에서 판매되는 쇠고기의 유전자 분석을 이용한 한우육 감별)

  • Kim Jin-Man;Nam Yong-Suk;Choi Ji-Hun;Lee Mi-Ae;Jeong Jong-Yon;Kim Cheon-Jei
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.203-209
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    • 2005
  • Real time-polymerase chain reaction (RT-PCR) is currently considered as the most sensitive method to detect low abundant DNAs in samples. Compared to conventional PCR, real-time PCR has a high reliability because of excluding false-positive results and can allow a simultaneous faster detection and quantification of target DNAs. This study was carried out to identify the Hanwoo (Korean native cattle) beef by genotyping after DNA extraction of commercial beef in 41 restaurants. Since Hanwoo, Holstein and imported cattle meat have different patterns in the MC1R gene associated with the coat colors of cattles (C-type, C/T-type or T-type), we could identify the genotype using real-time PCR The result of real-time PCR assay for beef samples in 41 restaurants which are asserted to sell Hanwoo beef only, showed that 29 of 41 samples were Hanwoo beef gene type (T-type) and 12 of 41 samples were Holstein or imported cattle gene type (C-type or C/T-type). Therefore, the proportion of Han-woo beef was $70.7\%$ and the proportion of Holstein or imported cattle meat was $29.3\%(C/T-type; 12.2\%,\;C-type; 17.1\%)$.

Identification of Irradiation -induced Volatile Marker Compounds in Irradiated Red Pepper Powder (방사선조사 고추가루로부터 휘발성 표지물질의 구명)

  • Kim, Hun;Ahn, Jun-Suck;Sin, Yeong-Min;Lee, Yong-Ja;Lee, Kyung-Hae;Byun, Myung-Woo;Cha, Yong-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.236-242
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    • 2005
  • To develop a new detection method using irradiation-induced volatile marker compounds of red pepper powder (RP), the volatile compounds of irradiated RP (0, 1, 3, 5, and 10 kGy) were analyzed by purge and trap (P&T)/solid phase microextraction (SPME)/gas chromatography/mass spectrometry (GC/MS) methods. A total of 51 and 31 compounds were detected in IRP by SPME and P&T methods, respectively. Among these, 25 compounds, which were composed of 4 hydrocarbons, 7 aldehydes, 1 ketone, 3 alcohols, 4 aromatic compounds, 2 esters and 4 miscellaneous compounds, showed irradiation dependent manner with significant positive correlation (p<0.01 or p<0.05) between irradiation dose and relative concentration. However, all compounds except 1,3-bis(1,1-dimethylethyl)benzene were not suitable as marker compounds because of their low determination coefficients ($R^2$<0.80) between irradiation dose and their concentrations, and detectablilty in nonirradiated sample. Therefore, only one compound, 1,3-bis(1,1-dimethylethyl)benzene, was tentatively identified as a volatile marker compound to detect irradiated RP.

Identification of a Single Nucleotide Polymorphism (SNP) Marker for the Detection of Enhanced Honey Production in Hoenybee (수밀력 우수 꿀벌 계통 판별을 위한 계통 특이 분자마커 개발)

  • Kim, Hye-Kyung;Lee, Myeong-Lyeol;Lee, Man-Young;Choi, Yong-Soo;Kim, Dongwon;Kang, Ah Rang
    • Journal of Apiculture
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    • v.32 no.3
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    • pp.147-154
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    • 2017
  • Honeybees (Apis mellifera) are common pollinators and important insects studied in agriculture, ecology and basic research. Recently, RDA (Rural Development Administration) and YIRI (Yecheon-gun Industrial Insect Research Institute) have been breeding a triple crossbred honey bee named Jangwon, which have the ability to produce superior quality honey. In this study, we identified a single nucleotide polymorphism (SNP) marker in the genome of Jangwon honeybee, particularly, in the paternal line (D line). Initially, we performed Sequence-Based Genotyping (SBG) using the Illumina Hiseq 2500 in 5 honeybee inbred lines; A, C, D, E, and F; and obtained 1,029 SNPs. Seventeen SNPs for each inbred line were generated and selected after further filtering of the SNP dataset. The 17 SNP markers validated by performing TaqMan probe-based real-time PCR and genotyping analysis was conducted. Genotyping analysis of the 5 honeybee inbred lines and one hybrid line, $D{\times}F$, revealed that one set of SNP marker, AmD9, precisely discriminated the inbred line D from the others. Our results suggest that the identified SNP marker, AmD9, is successful in distinguishing the inbred honeybee lines D, and can be directly used for genotyping and breeding applications.

Training Performance Analysis of Semantic Segmentation Deep Learning Model by Progressive Combining Multi-modal Spatial Information Datasets (다중 공간정보 데이터의 점진적 조합에 의한 의미적 분류 딥러닝 모델 학습 성능 분석)

  • Lee, Dae-Geon;Shin, Young-Ha;Lee, Dong-Cheon
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.40 no.2
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    • pp.91-108
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    • 2022
  • In most cases, optical images have been used as training data of DL (Deep Learning) models for object detection, recognition, identification, classification, semantic segmentation, and instance segmentation. However, properties of 3D objects in the real-world could not be fully explored with 2D images. One of the major sources of the 3D geospatial information is DSM (Digital Surface Model). In this matter, characteristic information derived from DSM would be effective to analyze 3D terrain features. Especially, man-made objects such as buildings having geometrically unique shape could be described by geometric elements that are obtained from 3D geospatial data. The background and motivation of this paper were drawn from concept of the intrinsic image that is involved in high-level visual information processing. This paper aims to extract buildings after classifying terrain features by training DL model with DSM-derived information including slope, aspect, and SRI (Shaded Relief Image). The experiments were carried out using DSM and label dataset provided by ISPRS (International Society for Photogrammetry and Remote Sensing) for CNN-based SegNet model. In particular, experiments focus on combining multi-source information to improve training performance and synergistic effect of the DL model. The results demonstrate that buildings were effectively classified and extracted by the proposed approach.

Epidemiology and Clinical Characteristics of Enterovirus Infections in Children: A Single Center Analysis from 2006 to 2010 (2006-2010년 단일기관 소아에서의 엔테로바이러스 감염 역학 및 임상 양상에 관한 연구)

  • Park, In-Soo;Lee, Hae Sung;Choi, Soo-Han;Kim, Hye Jin;Hwang, Seo Yeon;Cheon, Doo-Sung;Chang, Jin-Keun
    • Pediatric Infection and Vaccine
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    • v.20 no.2
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    • pp.81-88
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    • 2013
  • Purpose : This study was performed to investigate the epidemiology of enterovirus (EV) infections in children at a secondary hospital during recent 5 years. Methods : We collected the cerebrospinal fluid, stool and throat swab samples from the pediatric patients with suspected EV infections in KEPCO Medical Center, Seoul, Korea from July 2006 to September 2010. EV detection and genotype identification were performed by RT-PCR at Korea Centers for Disease Control and Prevention. Results : A total of 386 samples were collected from 277 patients during study period. Ninety-eight patients (35.4%) were diagnosed with EV infections. The RT-PCR positive rate was the highest in throat swab samples (48.3%). The median age of patient was 4.7 years (range, 0.1-12.5 years). Aseptic meningitis (50, 51.0%) was the most common clinical manifestation; herpangina (22, 22.4%) and hand-foot-mouth disease (18, 18.4%). One hundred EVs were isolated from 98 patients and 20 genotypes of EV were identified; Echovirus 30 (28 cases, 28%), Enterovirus 71 (12 cases, 12%), Echovirus 25 (10 cases, 10%), Echovirus 9 (9 cases, 9%) and Coxsackievirus A6 (8 cases, 8%). Aseptic meningitis caused by Echovirus 30 was the most common manifestation in 2008. There was no complicated case caused by Enterovirus 71. Conclusion : This study showed the epidemiology of confirmed EV infection in children from 2006 to 2010. There is a need for continuous surveillance of EV infections and its clinical manifestations.

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Detection and Identification of Mycobacterium Tuberculosis in Patients with Tuberculous Cervical Lymphadenitis by PCR-RFLP (경부 결핵성 임파선염 환자에서 PCR-RELP를 이용한 결핵균의 검출 및 확인)

  • Lee Sang-Sook;Cho Young-Rok;Chun Ji-Min;Choi Yong-Seok;Sohn Eun-Ju;Park Nam-Cho;Park June-Sik
    • Korean Journal of Head & Neck Oncology
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    • v.12 no.2
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    • pp.169-176
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    • 1996
  • Tuberculous cervical lymphadenitis is still an important cause of neck mass in Korea. Tuberculosis is an important differential diagnosis in patients of cervical lymphadenopathy. Rapid and sensitive test for the diagnosis of tuberculosis is essential for the approapiate treatment. Up to now, conventional diagnostic methods for M. tuberculosis were acid-fast bacilli(AFB) stain and culture of M. tuberculosis. The direct microscopic examination of AFB by Ziehl-Neelsen stain is rapid, but often negative. The culture for M. tuberculosis is time-consuming, taking 4 to 8 weeks. Recently various methods to detect Mycobacterial DNA, including PCR and restriction fragment length polymorphism(RFLP) analysis have been reported. Here we represent a simple method for the confirmation of M. tuberculosis and exclusion of the other Mycobacterial species by RFLP analysis and silver staining of polyacrylamide gel electrophoresis after nested PCR for a repetitive DNA sequence(IS986) specific for M. tuberculosis from fresh or paraffin-embedded biopsy specimens. This result leads us to conclude that this method is simple, rapid and possibly applicable to confirm M. tuberculosis and rule out the other Mycobacteria species from the clinical specimens in the clinical laboratories.

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Rapid Drug Susceptibility Testing for Isoniazid and Rifampicin by Reverse Hybridization Assay (역교잡반응법을 이용한 아이소니아지드 및 리팜피신 신속감수성검사)

  • Park, Young Kil;Yu, Hee Kyoung;Ryu, Sung Weon;Bai, Gill Han
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.440-448
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    • 2003
  • Background : Development of rapid drug susceptibility testing provides the opportunity for rapid identification of individuals with drug resistant tubercle bacilli, allowing selection of appropriate therapeutic regimens. Methods : A total of 502 drug resistant isolates were subjected to reverse blot hybridization assay to detect mutations within genes (rpoB, katG, inhA, and ahpC) associated with rifampicin (RMP) and isoniazid (INH) resistance. Results : Among the 264 RMP resistant strains ($RMP^R$) tested, the most prevalent mutation was the Ser531Leu seen in 121 strains (46%). The second common mutation occurred in 84 strains (32%) at codon 526. And 27 strains (10%) showed the mutation at codon 516. Among all 469 INH resistant strains ($INH^R$), the katG mutation was responsible for INH. The inhA mutation was present in 88 strains (19%). In 11 isolates (2%), coexisting of the katG and inhA mutations were identified. Reverse hybridization assay successfully detected over 80% of $INH^R$ and over 92% of $RMP^R$ among Korean isolates. CONCLUSION: Reverse hybridization was useful for rapid detection of $INH^R$ and $RMP^R$.

Detection of Gastric Contraction in Electrogastrography: Spectrum Analysis and Vector Analysis (위전도에서의 위수축 측정방법 : 주파수영역분석 및 벡터분석)

  • Kim, In-Young;Han, Wan-Taek
    • Journal of Biomedical Engineering Research
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    • v.18 no.3
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    • pp.273-283
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    • 1997
  • Electrogastrography(EGG), the cutaneous recording of the myoelectrical activity of the stomach using surface electrodes, is attactive due to its non-invasiveness. Despite many attempts made over the decades, analysis of surface EGG has not led to identification of contraction-related electrical response activity of the stomach that would help the clinician to diagnose motility disorders of the stomach. We propose feasible methods to detect the gastric contraction by spectrum analysis and vector analysis of the surface EGG signal. A running spectral analysis(RSA) based on the fast Fourier transform (FFT) was applied to the filtered EGG signal. The powers of dominant frequency and its harmonics were compared with gastric contraction signals such as the strain gauge signal from the gastric serosa in dog or the antropyloric pressure in human. And we also carried out vector analysis of the filtered EGG signals obtained from three paired electrodes. The amplitude and direction of the calculated EGG vector were analyzed and compared with the gastric contraction signals. From the spectrum analysis, we found that the increase of the power of the first harmonic of the dominant frequency was highly correlated with the gastric contraction. And from the vector analysis of the EGG signal, we found a typical change of the amplitude and direction of the EGG vector, which can indicate occurrences of the gastric contraction.

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