• Title/Summary/Keyword: immunochromatographic assay(ICA)

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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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Comparison of an Immunochromatographic Assay Kit with DAS-ELISA for Large-Scale Diagnosis and Molecular Discrimination of Satsuma Dwarf Virus Collected from Citrus Orchards

  • Kato, Mitsuhiro;Tomimura, Kenta;Ishii, Kanako
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.509-514
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    • 2020
  • Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

Development of a Novel Immunochromatographic Assay for Rapid Detection of VanA Ligase-Producing Vancomycin-Resistant Enterococci

  • Ji, Gil Yong;Song, Hyung Geun;Son, Bo Ra;Hong, Seung Bok;Kim, Jong Wan;Shin, Kyeong Seob
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.427-430
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    • 2014
  • We developed a novel immunochromatographic assay (ICA) (EZ-Step VanA rapid kit; Dinona, Korea) for the detection of VanA ligase from vancomycin-resistant enterococci (VRE). Of eight monoclonal antibodies screened by ELISAs, the VanA ligase ICA constructed with 1H9 plus 3G11 showed the greatest reactivity. The detection limit of the kit was $6.3{\times}10^6$ CFU per test. Of 127 vancomycin-resistant microorganisms, 100 vanA VRE were positive in the VanA ligase ICA, and 27 non-vanA vancomycin-resistant isolates were negative. These results were consistent with those of the PCR analyses. Thus, our ICA is a reliable and easy-to-use immunological assay for detecting VanA-producing VRE in clinical laboratories.

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.

Quantitative Method of Rapid Immunochromatographic Assay Kit for HBsAg-screening using Computer Image Analysis (컴퓨터 상 분석을 이용한 HBsAg-screening용 Rapid Immunochromatographic Assay Kit의 정량적 측정법)

  • 신형순;허태련
    • KSBB Journal
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    • v.15 no.3
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    • pp.243-246
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    • 2000
  • One of recent topics in the case of hepatits B virus(HVB) is the value of hepatitis B surface antigen(HBsAg) concentration as a prognostic maker. We developed uantitative method of rapid immunochromatographic assay(ICA) kit for HBsAg using computer image analysis (CIA) for the purpose of home diagnosis. uantitative ICA using CIA demonstrated integrated optical density(IOD) values proportional to log of reference HBsAg concentrations in the range of 2-200 ng/mL and enzyme-linked immunosorbent assay(ELISA) demonstrated the same in the range of 0.1-100 ng/mL however the test results with sample sear showed the same concentration on both kits. Furthermore repeated tests with the same samples revealed that this quantitative ICA using CIA would be reproducible and coefficient of variation(CV) of the results was 1.38~6.30%.

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Development of Diagnostic kit for Hepatitis B Susrface Antigen using Immunochromatographic Assay Method (면역크로마토그래피법을 이용한 B형간염 진단용 kit의 개발)

  • 신형순;신광순;정홍근;허태련
    • KSBB Journal
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    • v.15 no.2
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    • pp.214-218
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    • 2000
  • A hepatitis B Surface Antigen(HBsAg)-screening kit using immunochromatographic assay(ICA) method was developed by e employing two kinds of antibodies. One is mouse monoclonal anti-HBs for tracer antibody and the other is goat p이yclonal a anti-HBs for capture antibody. This capture antibody was immobilized on the surface of nitroceliulose(NC) membrane and the t tracer antibody was conjugated with g미d particles. When serum sample was added to the sample well, the $\infty$njugates d deposited in a dry state on the surface of glass fiber filter were reconstituted and then combined with HBsAg in serum. In 5 5 min after adding, the assay result was visible through the window, that is, the complexes composed of HBsAg and the c conjugates appeared as maroon line on the lower part of the NC membrane. The detection limit of the ICA kit was 2 ng/ml w when being tested with the reference HBsAg.

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Comparison of Methods for the Detection of Anti-HBs for Hepatitis B Vaccination Program in Korea (보건예방사업을 위한 B형간염 표면항체 검사방법 비교)

  • Lee, Jeong-Nyeo;Urm, Sang-Hwa;Lee, Jong-Tae;Chun, Jin-Ho;Sohn, Hae-Sook
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.2
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    • pp.226-230
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    • 2000
  • Objectives : The purpose of this study was to suggest a proper method for the detection of heaptitis B surface antibody(anti-HBs) in a screening program for hepatitis B vaccination. Methods : Sensivitity, specificity and predictive values were compared between Immunochromatographic assay (ICA) and passive hemagglutination(PHA) in 978 subjects(565 males, 413 females, 19-78 years ranging in age, mean 46.5 years old). EIA was used as a standard method for the detection of HBsAb. Results : Sensitivity in the detection of anti-HBs of PHA and ICA was 88.7%, and 94.9%, specificity was 94.3% and 96.6%, negative predictive value was 96.5%, and 98.0%, and positive predictive value was 82.3%, and 91.3%,, respectively. False negative rate(11.3%) of PHA was higher than that(5.1%) of ICA. The higher the titer of anti-HBs in EIA was, the lower the false negative rate was. There was no false negative result in the cases with $101mIU/{\beta}c$ or more in EIA Conclusion : We suggest that ICA should be the choice of screening method in the detection of anti-HBs in Hepatitis B vaccination program.

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Usefulness of the BACTEC MGIT 960 System for Mycobacterial Culture and TB Ag MPT64 Immunochromatographic Assay to Identify Mycobacterium tuberculosis (항산균 배양에서 BACTEC MGIT 960 System의 유용성 및 MGIT 양성배지에서 결핵균 진단을 위한 TB Ag MPT64 면역발색법의 유용성)

  • Lee, Seung Hun;Lee, Min Jeong;Lee, Jeong-Mi;Yim, Su Jin;Lee, Seung Jun;Kim, You Eun;Cho, Yu Ji;Jeong, Yi Yeong;Kim, Ho Cheol;Lee, Jong Deog;Kim, Sun Joo;Hwang, Young Sil
    • Journal of Yeungnam Medical Science
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    • v.29 no.2
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    • pp.83-88
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    • 2012
  • Background: This study was conducted to evaluate the usefulness of the BACTEC MGIT (Mycobacterium Growth Indicator Tube) 960 system for mycobacteria culture and immunochromatographic assay to identify Mycobacterium tuberculosis (MTB) in positive MGIT culture. Methods: Mycobacteria-culture-positive cases were retrospectively analyzed from December 2010 to July 2011. The detection rates and the recovery times of the mycobacteria between the Ogawa media and the MGIT were compared. An immunochromatographic assay (ICA) (SD BIO-LINE) was also performed in the positive MGIT culture for identification, and the results were compared with those of the Ogawa media in the Korea National Tuberculosis Association. Results: Among the 261 patients (M:F, 168:93; mean age, $61.6{\pm}17.16$ yrs), 450 specimens (sputa, 365; bronchial washing, 61; and pleural effusion, 24) were found positive with mycobacteria. Mycobacteria were grown both on the MGIT and Ogawa media in 310 cases (68.9%); only on the MGIT in 115 cases (22.6%); and only on the Ogawa media in 25 cases (5.5%) (p<0.05). The recovery time was $28.2{\pm}8.9$ days in the Ogawa media and $11.1{\pm}5.8$ days in the MGIT (p<0.05). Among the 127 cases from the positive MGIT culture, all 92 cases that were confirmed as MTB cases bythe Korea National Tuberculosis Association were identified as MTB by ICA, with 100% sensitivity. Conclusion: MGIT increases the detection rate and shortens the recovery time of mycobacteria in clinical respiratory specimens, and the TB Ag MPT64 kit using ICA is useful in identifying MTB in a positive MGIT culture.

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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.

Sputum Processing Method for Lateral Flow Immunochromatographic Assays to Detect Coronaviruses

  • Aram Kang;Minjoo Yeom;Hyekwon Kim;Sun-Woo Yoon;Dae-Gwin Jeong;Hyong-Joon Moon;Kwang-Soo Lyoo;Woonsung Na;Daesub Song
    • IMMUNE NETWORK
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    • v.21 no.1
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    • pp.11.1-11.10
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    • 2021
  • Coronavirus causes an infectious disease in various species and crosses the species barriers leading to the outbreak of zoonotic diseases. Due to the respiratory diseases are mainly caused in humans and viruses are replicated and excreted through the respiratory tract, the nasal fluid and sputum are mainly used for diagnosis. Early diagnosis of coronavirus plays an important role in preventing its spread and is essential for quarantine policies. For rapid decision and prompt triage of infected host, the immunochromatographic assay (ICA) has been widely used for point of care testing. However, when the ICA is applied to an expectorated sputum in which antigens are present, the viscosity of sputum interferes with the migration of the antigens on the test strip. To overcome this limitation, it is necessary to use a mucolytic agent without affecting the antigens. In this study, we combined known mucolytic agents to lower the viscosity of sputum and applied that to alpha and beta coronavirus, porcine epidemic diarrhea virus (PEDV) and Middle East respiratory syndrome coronavirus (MERS-CoV), respectively, spiked in sputum to find optimal pretreatment conditions. The pretreatment method using tris(2-carboxyethyl)phosphine (TCEP) and BSA was suitable for ICA diagnosis of sputum samples spiked with PEDV and MERS-CoV. This sensitive assay for the detection of coronavirus in sputum provides an useful information for the diagnosis of pathogen in low respiratory tract.