• Title/Summary/Keyword: detection kit

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Application of the Onsite EEW Technology Using the P-Wave of Seismic Records in Korea (국내 지진관측기록의 P파를 이용한 지진현장경보기술 적용)

  • Lee, HoJun;Jeon, Inchan;Seo, JeongBeom;Lee, JinKoo
    • Journal of the Society of Disaster Information
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    • v.16 no.1
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    • pp.133-143
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    • 2020
  • Purpose: This study aims to derive a predictive empirical equation for PGV prediction from P-wave using earthquake records in Korea and to verify the reliability of Onsite EEW. Method: The noise of P wave is removed from the observations of 627 seismic events in Korea to derive an empirical equation with PGV on the base rock, and reliability of Onsite alarms is verified from comparing PGV's predictions and observations through simulation using the empirical equation. Result: P-waves were extracted using the Filter Picker from earthquake observation records that eliminated noises, a linear regression with PGV was used to derive a predictive empirical equation for Onsite EEW. Through the on-site warning simulation we could get a success rate of 80% within the MMI±1 error range above MMI IV or higher. Conclusion: Through this study, the design feasibility and performance of Onsite EEWS using domestic earthquake records were verified. In order to increase validity, additional medium-sized seismic observations from abroad are required, the mis-detection of P waves is controlled, and the effect of seismic amplification on the surface is required.

Development and Evaluation of an Immunochromatographic Assay for Screening Listeria spp. in Pork and Milk

  • Kim, Seong-Hee;Kim, Jin-Young;Han, Woong;Jung, Byeong-Yeal;Chuong, Pham-Due;Joo, Hae-Jin;Ba, Hoa-Van;Son, Won-Geun;Jee, Young-Heun;Yoon, Byoung-Su;Lee, Yong-Soon;Lim, Yoon-Kyu
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.515-519
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    • 2007
  • Rapid immunochromatographic assay (ICA) kits were developed using flagella-specific monoclonal antibodies (MAbs) and rabbit polyclonal antibodies for screening Listeria spp. in food. The establishment of different formats, MAb 2B1 as capture antibody and MAb 7A3 or rabbit polyclonal antibodies as detector antibody, was compared. The 2 formats of the ICA kit were shown to have specific reactions with Listeria and no cross-reactivity with any of the non-Listeria including Escherichia coli O157:H7 and Salmonella enteritidis. The detection limits of the ICA kit using the combination of gold-labeled MAb 7A3 and MAb 2B1 showed $1{\times}10^5$ and $1{\times}10^6\;CFU/0.1\;mL$ at 22 and $30^{\circ}C$, respectively. The other format of the ICA kit using the combination of gold-labeled rabbit polyclonal antibodies and MAb 2B1 showed $1{\times}10^6\;CFU/0.1\;mL$ at $22^{\circ}C$ but weak signal at 30 culture. The format utilizing MAb was more sensitive than the one using polyclonal antibodies for capture antibody. Samples contaminated with L. monocytogenes 4b culture (9-10, 5-6, and 1-2 CFU/mL) on pork and pasteurized milk were confirmed as positive results. Current data suggests that this ICA kit is a rapid, simple and effective tool to screen for Listeria spp. in food.

Development of a Novel Immunochromatographic Assay for Rapid Detection of OXA-23 β-lactamase-producing Acinetobacter baumannii

  • Ji, Gil Young;Song, Hyung Geun;Jo, Mi Young;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
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    • v.22 no.2
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    • pp.29-36
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    • 2016
  • Among the several agents causing carbapenem resistance of Acinetobacter baumannii, the most common cause is OXA-23 ${\beta}$-lactamase, which is known to hydrolyze carbapenem. To effectively control dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB), development of both rapid and easy-to-use detection methods are required. The aim of this study is to develop a novel immunochromatographic assay (ICA) for rapid detection of OXA-23 ${\beta}$-lactamase. Of the seven monoclonal antibodies (mAbs) screened by ELISA, four mAbs (4G6, 4H6, 6G4, 9A4) exhibited high reactivity. Of these four specific antibodies, the combination of 6G4/4G6 showed the greatest reactivity and this combination of mAbs (6G4/4G6 mAbs) was used to develop the OXA-23 ${\beta}$-lactamase ICA. Of 102 A. baumannii isolates tested, the OXA-23 ${\beta}$-lactamase ICA results were consistent with PCR analysis except one false positive and one false negative isolate. The overall sensitivity and specificity were 98.36% and 97.56%, respectively. In conclusion, to the best of our knowledge, we have developed the first specific antibody set to detect OXA-23 ${\beta}$-lactamase using an ICA kit. This novel ICA can be used as a reliable and easy-to-use immunological assay for detection of OXA-23 ${\beta}$-lactamase producing CRAB in clinical laboratories.

Development of a biosensor from aptamers for detection of the porcine reproductive and respiratory syndrome virus

  • Kuitio, Chakpetch;Rasri, Natchaya;Kiriwan, Duangnapa;Unajak, Sasimanas;Choowongkomon, Kiattawee
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.79.1-79.12
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    • 2020
  • Background: Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential. Objectives: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig. Methods: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM). Results: Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 1010 particles. Conclusions: SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.

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.

Optimal RNA Extraction Methods and Development of Synthetic Clones for Seven Strawberry Viruses (딸기바이러스 진단을 위한 최적의 RNA 추출 방법 및 주요 7종 딸기바이 러스의 진단법 개발)

  • Kwon, Sun-Jung;Yoon, Ju-Yeon;Cho, In-Sook;Chung, Bong-Nam
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.170-178
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    • 2020
  • Most strawberry viruses exist relatively low titers in tissues, and strawberry tissues include high levels of contamination by polysaccharides and phenolic compounds. These traits make the efficiency of strawberry diagnosis difficult. In this study, we tested different commercially available kits and reagents to secure optimal RNA extraction methods to determine virus detection from strawberry leaves. Total RNA was isolated from leaves of strawberry mottle virus (SMoV)-infected strawberry cultivar 'Mihong'. The efficiency of total RNA for virus diagnosis was confirmed through SMoV detection by one-step or two-step reverse transcription and polymerase chain reaction (RT-PCR). Among those, the RNeasy plant RNA kit was best to isolate RNA and the isolated RNA was good enough for further applications. To ensure a reliable detection for strawberry viruses, synthetic diagnosis clones for major seven strawberry viruses such as strawberry mild yellow edge virus, SMoV, strawberry latent ring spot virus, strawberry crinkle virus, strawberry pallidosis associated virus, strawberry vein banding virus and strawberry necrotic spot virus have been constructed. Based on the synthetic genes in each clone, primer sets for seven strawberry viruses were designed and tested an RT-PCR condition through a simultaneous application of the same annealing temperature that allowed to achieve an efficient and convenient diagnosis.

Rapid Detection of Pathogens Associated with Dental Caries and Periodontitis by PCR Using a Modified DNA Extraction Method (PCR을 이용한 치아우식증 및 치주염 연관 병원체의 빠른 검출)

  • Kim, Jaehwan;Kim, Miah;Lee, Daewoo;Baik, Byeongju;Yang, Yeonmi;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.41 no.4
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    • pp.292-297
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    • 2014
  • DNA extraction is a prerequisite for the identification of pathogens in clinical samples. Commercial DNA extraction kits generally involve time-consuming and laborious multi-step procedures. In the present study, our modified DNA isolation method for saliva samples allows for the quick detection of pathogens associated with dental caries or periodontitis by PCR within 1 h. To release DNA from the bacteria, 1 min of boiling was adequate, and the resulting isolated DNA can be used many times and is suitable for long term storage of at least 13 months at $4^{\circ}C$, and even longer at $-20^{\circ}C$. In conclusion, our modified DNA extraction method is simple, rapid, and cost-effective, and suitable for preparing DNA from clinical samples for PCR for the rapid detection of oral pathogens from saliva.

Comparison of detecting methods and the relationship between tissue and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens (육계에서 조직과 혈액의 enrofloxacin 및 대사성 ciprofloxacin 잔류 상관관계 조사 및 검사방법간의 비교)

  • Sung, Myung-Suk;Kim, Mi-Suk;Seo, Hee-Jin;Bae, Dong-Rok;Hwang, Ji-Young;Kim, Soon-Tae;Cho, Jong-Suk;Park, Hong-Je;Hong, Sung-Hee;Kim, Gyung-Dong;Jang, Seong-Jun;Yun, Mun-Jo
    • Korean Journal of Veterinary Service
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    • v.36 no.4
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    • pp.311-320
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    • 2013
  • The purpose of this study was to evaluate detecting methods and the relationship between tissues and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens. Two groups of broiler chickens were administrated via the drinking water with $50{\mu}g/mL$ and $100{\mu}g/mL$ of enrofloxacin for 5 days, respectively. The concentration of enrofloxacin and metabolic ciprofloxacin in tissues (muscle and kidney) and blood were measured during administration period (for 5 days) and withdrawal period (for 12 days) by high performance liquid chromatography (HPLC) method. Also, all samples were conducted for screening of residues by microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit. The relationship between tissues (muscle and kidney) and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens was followed : The levels of enrofloxacin and metabolic ciprofloxacin residues in muscle and kidney were higher 2.9~3.2 folds, 3.6~3.8 folds more than the residues levels in blood, respectively. These results support we can predict the residues in muscle and kidney from the residues in blood. In comparison of detecting methods for antibiotic residues, microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit could detect positive reaction at similar or lower concentration than violative concentration of enrofloxacin and metabolic ciprofloxacin in chicken tissues. These results support what two screening methods are useful for screening of quinolone detection in chickens.

Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods

  • Jang, Juno;Hong, Sung-Hwan;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.100-108
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    • 2011
  • A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 ($TCID_{50}$) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.

Application of the melanocortin 1 receptor (MC1R) gene for discrimination of Hanwoo from Holstein beef using real-time polymerase chain reaction (PCR)

  • Ra, Do-Kyung;Lee, Sung-Mo;Park, Eun-Jeong;Lee, Jung-Goo
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.557-562
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    • 2007
  • This study was carried out to discriminate Hanwoo from the milking and hybrid cattle by detection of MC1R gene related to bovine hair color. One hundred sixty six samples were collected from the abattoir (n = 106) and local market (n = 60). The beef from abattoir were originated from Hanwoo (n=27), Holstein (n=29), Hybrid (n=45) and imported cattle (n=5), respectively. The beef from market consisted of Hanwoo (n=36), Holstein (n=7) and imported ones (n=17). Commercialized screening kit (Kogenebiotec, Korea) was used for MC1R gene analysis. As a result, Hanwoo was discriminated from Holstein. However, 9 of 45 hybrid and 11 of 22 imported beef samples were indistinguishable from Hanwoo. It could be explained by second generation of crossing of Hanwoo with Holstein or the cattle with silver or yellow hair. This results suggest that additional tests as well as MC1R gene detection be needed to confirm Hanwoo beef among cattle beef.