• Title/Summary/Keyword: Kit

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Comparison of a PCR Kit and a Selective Medium to Detect Pathogenic Bacteria in Eggs (PCR Kit와 선택배지를 이용한 계란의 병원성세균 검출 비교 평가)

  • Kim, Dong-Ho;Yun, Hye-Jeong;Song, Hyun-Pa;Lim, Sang-Yong;Jo, Min-Ho;Jo, Cheo-Run
    • Food Science and Preservation
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    • v.16 no.6
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    • pp.965-970
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    • 2009
  • PCR technology has been widely used to detect and quantify microbial pathogens in foodstuffs, because the technique is rapid, sensitive, and selective. In this study, detection of contaminating pathogenic bacteria on shells of chicken eggs was performed using both a commercial multiplex polymerase chain reaction (PCR) kit and a viable count method employing a selective medium. The PCR kit was capable of detecting Campylobacter jejuni, Escherichia coli O157:H7, Staphylococcus aureus, Bacillus cereus, Vibrio parahaemolyticus, Listeria monocytogenes, Yersinia enterocolitica, Salmonella species, and Shigella species. Using the PCR method, five bacterial species were detected from 30 samples (33.3%) of 90 batches of eggs commercially available in a market. PCR products from B. cereus, S. aureus, L. monocytogenes, Y. enterocolitica, and E. coli O157:H7 were detected, and the numbers and frequencies of positive samples were 17 (18.8%), 12 (13.3%), 15 (16.6%), 16 (17.7%),and 4 (4.4%), respectively. None of any Salmonella species, C. jejuni, V. parahaemolyticus, or Shigella species was detected in this study. The results of PCR testing were confirmed using a typical viable count method employing a selective medium. We suggest that the multiplex polymerase chain reaction (mPCR) assay is a rapid and reliable method for detection of pathogenic bacteria contaminating eggs.

Development of Infectious Bronchitis Virus (IBV) ELISA Kit for Detection of Antibodies against Nephropathogenic IBV Vaccine (국내회사와 다국적기업 제조 ELISA 키트의 전염성 기관지염 백신에 따른 항체 검출능 비교)

  • Kim, Kyu-Jik;Kim, Jun-Young;Youn, Ha-Na;Ju, Hyo-Sun;Lee, Da-Yeah;Song, Chang-Seon
    • Korean Journal of Poultry Science
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    • v.45 no.1
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    • pp.17-28
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    • 2018
  • Infectious bronchitis virus (IBV) is an economically important disease in the poultry industry worldwide. This disease commonly manifests respiratory signs, poor egg quality, and decline in egg production. Since IBV is a RNA virus, the emergence of new variant strains is continuously reported and the immunization of susceptible chickens with only one antigenic type of the virus has been shown to induce partial or no protection against other unrelated types. Therefore, it is difficult to diagnose IBV due to variants serotypes. In this study, we collected serum from various ages of Broiler GP (Grandparent) to Layer CC (Commercial chick) and performed detectability comparison test between domestic company and multinational company manufacturing ELISA kit. Results of this experiment suggest that domestic company manufacturing ELISA kit is more sensitive to infectious bronchitis antibody than that of the multinational company. Our findings also suggest that antibody's change trends after infectious bronchitis vaccination. Thus, the use of appropriate kit for domestic situations is important.

Genetic Characterization of Wolla Coat Color in Jeju Horses (제주마에서 월라 모색의 유전적 특성)

  • Kim, Nam-Young;Shin, Kwang-Ynu;Lee, Chong-Eon;Han, Sang-Hyun;Lee, Sung-Soo;Park, Yong-Sang;Ko, Moon-Suck;Hong, Hyun-Ju;Yang, Jae-Hyuk;Jang, Deok-Jee;Yang, Young-Hoon
    • Journal of Animal Science and Technology
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    • v.54 no.5
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    • pp.375-379
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    • 2012
  • This study was carried out to define the "Wolla" coat color using 376 Jeju registered horses (white patched 142, solid coat color 234). Three major factors related to the white patches i.e ECA3-inversion for Tobiano, EDNRB 2 bp nucleotide substitution for frame Overo, and the KIT intron 16 single nucleotide polymorphism (SNP) for Sabino types of coat color were analyzed. It was found that out of 142 Jeju horses with white patches that have the genotype for ECA3-inversion (To) 140 horses were +/To heterozygous and 2 horses were To/To homozygous all Jeju horses with white patches had ECA3-inversion allele. However, there was no frame Overo or Sabino allele type in EDNRB and KIT intron 16 SNP in Jeju horses with white patches. As for 234 Jeju horses with a solid coat color, there was no ECA3-inversion allele related to the white patches. Thus, it could be considered that Wolla coat color with white patches in Jeju horses might have come from the Tobiano line in the genetic classification by coat color.

Development of Test System for Detection of Antibody to Human Immunodeficiency Virus Type 1 Subtype O (HIV-1 O형 항체 진단시료의 개발)

  • Cho, Young-Shik;Yu, Seung-Shin;Ha, Gun-Woo;Lee, Sang-Gook;Cho, Myung-Hwan;Shin, Hyung-Sik;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.31-38
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    • 1998
  • In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.

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Development of Enzyme Immuno Assay for Analysis of Free Prostate Specific Antigen in Serum (혈청 유리형 전립선항원 (free PSA) 측정을 위한 효소면역측정법의 개발)

  • Kyung-Ok Lee;Kyung-In Kim;Kyu-Pum Lee
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.107-114
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    • 1997
  • Recent reports indicate that the clinical usefulness of prostate specific antigen (PSA), particulary in the differentiation of benign prostate hyperplasia from prostate cancer, can be improved by measuring the amount of free PSA in serum. Measuring free PSA is especially useful in attempts to improve diagnositc performance of PSA in the diagnostic gray zone of total PSA. The objective of this study was to develop free PSA assay kit using sandwich microplate enzyme immunoassay format. We chose a test format with polyclonal anti-PSA antibodies coated on the wells and monoclonal anti-free PSA antibodies for quantification to gain higher test sensitivity. We adpoted 10 uL of specimen and 2 hours of first incubation time with detecting antibody for free PSA EIA format using microplate. The within-day and between-day precision (%CV) in the high and low concentration ranges were below 4%. The correlation coefficient between in-house free PSA assay and commercial assay kit was r=0.9965 (slope=0.0984, y intercept=0.0173, N=27). No hook effect was found by 40 ng/mL and correlation coefficient (r) value of the fitted linear regression was over 0.995. The recovery tests were in the range of 98.9∼104.1% for free PSA. In conclusion, in-house free PSA enzyme immune assay is cost effective, simple and rapid and could be useful for the prognosis after theraphy as well as for the differential diagnosis between prostate cancer and benign prostate hyperplasia.

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Usefulness of the Neutrophil Gelatinase-Associated Lipocalin (NGAL) Kit for Acute Kidney Injury Patients at the Emergency Medical Center in Daegu (대구지역 응급의료센터에 내원한 급성 콩팥손상 환자의 진단을 위한 호중구 젤라티나제 관련 리포칼린 키트의 유용성)

  • Lee, Seung-Jin;Park, Sangwook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.49-53
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    • 2016
  • Acute kidney injury (AKI) is a common syndrome resulting in kidney damage and malfunction within a few days or even a few hours. The diagnosis of AKI depends on routine biochemical tests, including serum creatinine, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), blood urea nitrogen (BUN), and electrolytes. Plasma neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker that shows correlation with the severity of acute infections and kidney injuries. The predictive value in other conventional assays for kidney functions has been reported to cause distraction for AKI syndrome. The aim of this study is to verify the predictive value of plasma NGAL in patients with established AKI. The NGAL kit for checkup demonstrates sensitivity of ${\geq}300$ (92.2%), ${\geq}200$ (95.6%), ${\geq}100$ (99.6%), specificity of ${\geq}300$ (95.1%), ${\geq}200$ (97.3%), ${\geq}100$ (99.4%), positive predictability of ${\geq}300$ (93.3%), ${\geq}200$ (93.4%), ${\geq}100$ (99.2%), and negative predictability of ${\geq}300$ (96.7%), ${\geq}200$ (97.7%), ${\geq}100$ (98.1%), respectively. The plasma NGAL compared with the enzyme-linked immunosorbent assay (ELISA) has been shown to be an early predictive biomarker of AKI. The NGAL kit, recently developed for point-of-care of plasma specimens, is thought to be a useful and reliable biomarker for the early diagnosis of decreased kidney functions.

Application of Breed-specific DNA Markers for the use of Identifying Major Pure Pig Breeds Maintained in Korea (대한민국내 주요 돼지 품종의 순종 식별을 위한 품종특이 DNA marker의 활용)

  • Seo, B.Y.;Kim, J.H.;Park, E.W.;Lim, H.T.;Cho, I.C.;Kim, B.W.;Oh, S.J.;Cheong, I.C.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.735-742
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    • 2004
  • This study was conducted for the identification of pure Landrace, Large White and Duroc breeds which are mainly maintained in Korea using DNA markers. We used known KIT and MC1R mutations, which were related coat color in pigs, and pig mitochondrial DNA variations. The KIT mutation was used to distinguish white and colored animals. Duroc breed could be discriminated from other colored breeds using the MC1R mutation N121D. Discriminating Landrace and Large White was possible using the l l-bp duplication of D-Ioop region and alternative initiation codon of ND2. In conclusion, identification of Landrace, Large White and Duroc breeds was might be possible using the procedure designed in this study.

Establishment of Complex Disaster Scenario on the Utility Tunnel Study for Digital Twin System Application (디지털트윈 시스템 적용을 위한 공동구 복합재난 시나리오 구축)

  • Yon Ha Chung; So Dam Kim;Hyun Jeong Seo;Hojun Lee;Tae Jung Song
    • Journal of the Society of Disaster Information
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    • v.18 no.4
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    • pp.861-872
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    • 2022
  • The purpose of this study was to establish a complex disaster scenario that can comprehensively consider various disaster situations that may occur in the utility tunnel. Method: In order to comprehensively consider the correlation between disasters, a composite disaster scenario was derived from a combination of damage factors, respectively. A risk assessment was performed in order to derive the priorities of the scenarios. And based on the results, the priorities of complex disaster scenarios were set. Result: Based on the disaster cases in the utility tunnel, a plan was prepared for complex disaster scenarios centered on damage. A complex disaster scenario was specified using a semi-quantitative evaluation method for single and multiple disaster factors such as fire, flooding, and earthquake. Conclusion: The composite disaster scenario derived from this study can be used for the prevention and preparation of damage when the precursor symptoms of a disaster are detected. In addition, the results of this study are expected to be used as basic data for preparing strategic plans and preparing complex disaster response technologies to induce rapid response and recovery in case of emergency disasters.

Efficiency Comparison of Environmental DNA Metabarcoding of Freshwater Fishes according to Filters, Extraction Kits, Primer Sets and PCR Methods (분석조건별 담수어류의 환경 DNA 메타바코딩 효율 비교: 필터, 추출 키트, 프라이머 조합 및 PCR 방법)

  • Kim, Keun-Sik;Kim, Keun-Yong;Yoon, Ju-Duk
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.199-208
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    • 2021
  • Environmental DNA (eDNA) metabarcoding is effective method with high detection sensitivity for evaluating fish biodiversity and detecting endangered fish from natural water samples. We compared the richness of operational taxonomic units(OTUs) and composition of freshwater fishes according to filters(cellulose nitrate filter vs. glass fiber filter), extraction kits(DNeasy2® Blood & Tissue Kit vs. DNeasy2® PowerWater Kit), primer sets (12S rDNA vs. 16S rDNA), and PCR methods (conventional PCR vs. touchdown PCR) to determine the optimal conditions for metabarcoding analysis of Korean freshwater fish. The glass fiber filter and DNeasy2® Blood & Tissue Kit combination showed the highest number of freshwater fish OTUs in both 12S and 16S rDNA. Among the four types, the primer sets only showed statistically significant difference in the average number of OTUs in class Actinopterygii (non-parametric Wilcoxon signed ranks test, p=0.005). However, there was no difference in the average number of OTUs in freshwater fish. The species composition also showed significant difference according to primer sets (PERMANOVA, Pseudo-F=6.9489, p=0.006), but no differences were observed in the other three types. The non-metric multidimensional scaling (NMDS) results revealed that species composition clustered together according to primer sets based on similarity of 65%; 16S rDNA primer set was mainly attributed to endangered species such as Microphysogobio koreensis and Pseudogobio brevicorpus. In contrast, the 12S rDNA primer set was mainly attributed to common species such as Zacco platypus and Coreoperca herzi. This study provides essential information on species diversity analysis using metabarcoding for environmental water samples obtained from rivers in Korea.