• Title/Summary/Keyword: detection kit

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Development of cellulose nano beads based a rapid detection kit to detect staphylococcal enterotoxin B

  • Kim, Giyoung;Yoo, Jinyoung;Park, Saetbyeol
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.549-557
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    • 2019
  • Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme-linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.

Effects of Optimal Heat Detection Kit on Fertility after Artificial Insemination (AI) in Hanwoo (Korean Native cattle) (한우 인공수정에서 수정적기 진단키트 활용이 수태율에 미치는 영향)

  • Choi, Sun-Ho;Jin, Hyun-Ju
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.153-157
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    • 2017
  • This study was conducted to investigate the optimal artificial insemination (AI) time with diagnostic kit at ovulation time. We already applied the patent about the protein in the cow heat mucose in external reproductive tract. And we would examine the accuracy for detection of cow heat by the kit produced with the protein. Evaluation of optimal heat detection was tried two time at 12 hrs and 24 hrs after the heat. And then, AI service also performed two times with no relation to the results of heat diagnosis by heat detection kit and pregnancy rates were checked with rectal palpation on $60^{th}$ day after AI. Heat diagnostic results by kit in natural heat after 12 hrs in Hanwoo cows were showed 31.3~75.0% on positive in first heat detection and 33.3~100.0% on positve in second heat detection. In the $1^{st}$ positive results were significant different (p<0.05), but $2^{nd}$ positive were not. The results of heat detection showed different result on regional influence and individual cow effects. The pregnancy rates of first trial of heat detection were showed 34.4~78.7% on positive and 21.3~68.8% on negative after the diagnosis by heat detection kit. And the pregnancy rates of next trial of heat detection were showed 33.3~85.7% on positive and 14.3~66.6% on negative after the heat diagnosis. Both positive results of first trial and next trial also were showed significant different (p<0.05), but negative results were not. In positive result, first trial of total pregnancy rates was higher than the next trial of pregnancy, but there showed opposite results on negative results. In conclusion, the optimal heat detection kit is suitable to ordinary Hanwoo cows and it suggested that we have to improve the kit's accuracy by detecting the materials like proteins related optimal AI time.

Development and Validation Study of Biological Agent Detection Kit (생물학작용제 검출 키트 개발 및 성능시험 연구)

  • Joe, Hae Eun
    • Journal of the Korea Institute of Military Science and Technology
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    • v.22 no.4
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    • pp.575-580
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    • 2019
  • In biological warfare, it is important to identify biological agents for proper treatment. We focused on developing a real-time RT-PCR kit that can detect multiple species of biological agents. AccuPower(R) Biothreat Real-Time RT-PCR Kit(v3.0) could detect Bacillus anthracis, Yersinia pestis, Vibrio cholerae, Francisella tularensis, Salmonella typhi, Rickettsia prowazekii, Variola virus, Hantaan virus, Yellow fever virus, Brucella spp., Shigella dysenteriae in a single reaction. The results showed that the kit was verified to be able to detect at least 0.005 ng of nucleotide and 10,000 CFU/ml of bacteria. Therefore, the kit is expected to be used as a rapid and sensitive detection kit for 11 species of biological agents within 2 hours.

Development of Multi-Residue Methods for Carbamate Pesticides by the Enzyme Inhibition Test (효소 저해법을 이용한 Carbamate계 농약의 다성분 잔류분석법 개발)

  • Kim, Jung-Ho
    • Journal of Environmental Science International
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    • v.17 no.12
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    • pp.1325-1330
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    • 2008
  • This study was carried out with the detection for multiresidue of the carbamate pesticide such as carbaryl and cabofuran by enzyme-inhibition method. The check time for determination of acetylcholinesterase(AChE) activity was selected at 60 sec. The AChE activity in chicken brain determined by the Ellman's method was $162{\mu}$mol/min/g protein. $I_{50}$ for AChE by carbamate pesticide with wet kit was 0.169mg/L of carbaryl and 0.089mg/L of cabofuran, respectively. The incubation time for enzyme kit with substrate kit was 30min for determination of AChE activity. Enzyme kit with substrate kit was stable at $4^{\circ}C\;and\;25^{\circ}C$ for 5 days. Limit detection concentration of carbaryl with dry kit for AChE was 0.05mg/L. The dry kit such as wet kit applied Enzyme-Inhibition(EI) method with AChE was confirmed the multi residue method to detect the carbamate pesticides.

Performance Evaluation of PCR Kits for Detecting Genetically Modified Crop Ingredients (유전자 변형 작물 성분 검출용 PCR Kit의 성능 평가 연구)

  • 윤시온;정순천;윤원기;박상규;문제선;이정현;김환묵
    • Toxicological Research
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    • v.20 no.2
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    • pp.101-108
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    • 2004
  • The different social reflections about the benefits and the potential risks of genetically modified (GM) crops have evolved with .different reactions in different countries. Many countries including Korea are working toward setting down new guidelines. Korea requires companies to label all food that contains more than 3% GM ingredients. One of the rapid and convenient detection methods of GM ingredients is amplification of the introduced DNAs by polymerase chain reaction (PCR). Many PCR kits for this purpose are commercially available. The objective of this study was to evaluate performance of commercialized GM crop detection kits. The results showed that 6 out of 15 kits tested did not meet the requirements even purposed by the manufacturers themselves in terms of stability, reproducibility, and detection limits, suggesting a potential quality control problem in their design stage or production line. The evaluation also suggests that, although the duplex and triplex detection kits allowed unambiguous detection in a single PCR reaction, the monoplex detection kits were the most sensitive to the detection of GM ingredients. The detection limits also differ between soybean and corn. Results from this study will be useful in the development of sound qualitative tracking systems of GM ingredients for monitoring throughout the cultivation of GM crops, their trans-boundary movement, and food production using GM grains as well as for complying with government guidelines associated with GM crops.

The Evaluation of Immunochromatographic Assay kit for Rapid Detection of Hepatitis B Surface Antigen (Hepatitis B Surface Antigen을 신속히 검출하기 위한 Immunochromatographic Assay kit의 성능 평가)

  • Shin, Hyeong-Soon;Kim, Young-Bong;Shin, Jung-Woo;Kim, Chang-Kyu;Lee, Wang-Sik;Kim, Han-Kyeom;Shin, Kwang-Soon
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.137-141
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    • 1997
  • We evaluated Immunochromatographic assay kit to screen HBsAg in human serum. When the reference HBsAg was applyed to ICA, HA and EIA kits, the limit of detection for HBsAg were found out to be 4, 2 and 0.25 ng/ml respectively. But ICA kit required 5 minutes to read the result whereas HA and EIA kit more than one hour. The sensitivity was 97% (29 of 30 samples) and the specificity 100% (45 samples) compared with conventional EIA. The ICA kit needs no instrument or machine to perform the test contrary to the conventional methods. Therefore, this rapid and sensitive ICA kit can be used for HBsAg-screening, especially in the emergency room and in the scene of the accident.

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Evaluation of the Efficiency of E. coli O157: H7 Rapid Detection Kit using Immunochromatography (면역크로마토그래피를 이용한 E. coli O157: H7 신속검출 키트의 유효성 평가)

  • Kwak, Hyo-Sun;Lee, Dong-Ha;Moon, Hee-Sook;Park, Jong-Seok;Woo, Gun-Jo;Kim, Chang-Min
    • Journal of Food Hygiene and Safety
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    • v.18 no.3
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    • pp.118-124
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    • 2003
  • For the rapid detection of various pathogenic microorganisms from food sample, various kinds of kits have been developed and commercially available in the markets. With the advantages of speed, accuracy and easiness, the market of these kits has gradually increased for the QC and QA field of food company as well as testing facilities or laboratories. In this study, the characteristics such as the detection limit and the sensitivity of immunochromatographic type of rapid detection kit (Donga Co, Korea, D-kit) for E. coli 0157:H7 developed by monoclonal antibody were examined and also the possibility of application of the kit to food samples was evaluated. The reference kits used for comparison study were Reveal E. coli 0157:H7 (Neogen Co., USA, R-kit) and VIP EHEC kit (Biocontrol Inc., USA, V-kit) occupying major market share. In the detection limit test with the E. coli 0157:H7 reference, both R-kit and D-kit showed a distinct positive reaction in $10^4$/ml and weak positive reaction in $10^3$/ml, whereas V-kit showed a same reaction in 105/ml. Also, it was identified that the culture treated with heat showed more sensitivity than no heat treated culture. The sensitivity test was conducted against 22 isolates of E. coli 0157:H7, 7 strains of non-O157:H7 verotoxin-producing E. coli, 40 strains of E. coli with different O and H antigen type, and 38 strains of non-E. coli Enterobacteriaceae, and all of the test strains except three were showed exactly three were showed exactly the same reaction against three kinds of the tested kits. All the three kinds of kits showed a positive reaction against E. coli O157:H19, E. coli O148:H18 and Salmonella galinarium. We suppose that there might be a similarity in serological property between these three strains and O157:H7. From the test results, it can be concluded that there is (was) no difference between the D-kit developed in this study and R-kit or V-kit based on the detection limit and sensitivity.

Sensitivity study of the Yfiler® PLUS PCR Amplification Kit in forensic casework samples (법과학 현장시료에서 Yfiler® PLUS PCR Amplification Kit의 민감도 연구)

  • Jung, Ju Yeon;Kim, Kyoung Sook;Park, Sun Wha;Lim, Si Keun;Lee, Dong Sub;Lee, Yang Han
    • Analytical Science and Technology
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    • v.29 no.1
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    • pp.43-48
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    • 2016
  • A variety of Y-STR analysis kits have been developed and used in the forensic field. Prior to the forensic application of a new kit, laboratory validation and sensitivity tests are essential processes in selecting suitable alternatives and for assuring that standard operating procedures are followed. In this paper, we have performed a sensitivity study of a new commercial kit, the Yfiler® PLUS PCR Amplification Kit (Yfiler plus kit, released in 2014) by comparing it with the AmpF/STR® YfilerTM PCR Amplification Kit (Yfiler kit, released in 2004). The Yfiler plus kit includes the 17 Y-STR loci of the Yfiler kit and has been supplemented with 10 new Y-STR loci. First, we analyzed the sensitivity difference between the two kits using commercial control DNA 2800M and 007. In addition, we compared the detection rate between the two kits from the 16 selected forensic casework samples of less than 0.5 ng concentrations. The results show that the sensitivity and detection rate of the Yfiler plus kit are higher than the corresponding rates of the Yfiler kit. In addition, we were able to obtain more Y-STR profiles with the use of the new kit. Thus, we suggest that Yfiler plus kit is a more effective forensic tool to detect Y-STR profiles from forensic casework samples of low concentrations.

Detection of Methicillin-resistant Staphylococcs aureus from the Anterior Nares of Healthcare Workers in a Intensive Care Unit by Using PBP2a Rapid Kit and Direct Coagulase Test (중환자실에 근무하는 의료인의 전비강에서 PBP2a Rapid Kit와 직접 Coagulase 검사를 이용한 Methicillin-resistant Staphylococcus aureus의 검출)

  • Hong, Seung-Bok;Shin, Kyung-A;Son, Jae-Cheol;Shin, Seob-Kyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.2
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    • pp.86-91
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    • 2010
  • We evaluated the performance of a novel screening test, PBP2a MRSA rapid kit (Dinona Inc., Iksan, Korea), for methicillin-resistant Staphylococcus aureus (MRSA) based on a immunochromatographic assay. The test is able to detect penicillin-binding protein 2a (PBP2a) using the nasal specimens from health care workers. The nasal specimens were obtained from 69 healthcare workers and were incubated in enrichment broth followed eight hours incubatin in BHI with cefoxitin $4{\mu}g/mL$. These broth were tested by PBP2a Rapid Kit. The enrichment broths were also directly tested for tube coagulase using the conventional identification method. 19 of 22 MRSA showed positive results by PBP2a rapid test and direct coagulase test (the sensitivity for detection of MRSA, 86.36%). While, 8 of 47 non-MRSA showed false positive results for the two tests. All of the 8 non-MRSA which showed false positive were co-colonizing isolates with MRCNS and MSSA. In addition, 46 of 49 methicillin-resistant staphylococci (MRS) showed positive results for PBP2a MRSA rapid kit (the sensitivity for detection of MRS, 93.8%), and all of 20 non-MRS showed negative results (specificity, 100%). The combination of PBP2a MRSA rapid kit and direct coagulase test showed the good sensitivity for detection of MRSA from anterior nares but frequently showed false positive results from the co-colonizing carrier with MRCNS and MSSA.

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Clinical evaluation of a rapid diagnostic test kit for detection of canine coronavirus

  • Yoon, Seung-Jae;Seo, Kyoung-Won;Song, Kun-Ho
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.27-31
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    • 2018
  • Canine coronavirus is a single-stranded RNA virus that causes enteritis in dogs of any age. Coronaviral enteritis is seldom definitively diagnosed, since it is usually much less severe than many other types of enteritis and is self-limiting. Conventional diagnostics for the canine coronaviral enteritis such as polymerase chain reaction (PCR), virus isolation, and electron microscopic examination are inappropriate for small animal clinics due to the complicated experimental processes involved. Therefore, a commercially available lateral flow test kit based on chromatographic immunoassay techniques was tested to evaluate its performance as a first-line diagnostic test kit that could be used in clinics. The coronavirus antigen test kit detected canine coronavirus-infected dogs with 93.1% sensitivity and 97.5% specificity. The detection limit of the test kit was between $1.97{\times}10^4/mL$ and $9.85{\times}10^3/mL$ for samples with a 2-fold serial dilution from $1.25{\times}10^6\;TCID_{50}$ ($TCID_{50}$, 50% tissue culture infectious dose). Additionally, the test kit had no cross-reactivity with canine parvovirus, distemper virus, or Escherichia coli. Overall, the commercially available test kit showed good diagnostic performance in a clinical setting, with results similar to those from PCR, confirming their potential for convenient and accurate use in small animal clinics.