• Title/Summary/Keyword: antigen test kit

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Nonspecific Mouse Hepatitis Virus Positivity of Genetically Engineered Mice Determined by ELISA

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.9-14
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    • 2015
  • Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

Evaluation of Immunotoxicities of New Skin Decontamination Kit(SDK) (신규 개인제독키트, SDK시제품(가칭)의 면역독성평가에 관한 고찰)

  • 강병철;이학모;제정환;남정석;이석만;양재만;박재학;이영순
    • Toxicological Research
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    • v.14 no.2
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    • pp.205-209
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    • 1998
  • To evaluate immunotoxicity of skin decontamination kit(SDK) newly-developed in Agency for Defense Development(ADD), delayed contact hypersensitivity (maximization) test and passive cutaneous anaphylaxis(PCA) test of SDK were performed and the results were compared with those of M 291. In maximization test, sensitization reaction was induced by id injection (2.5 mg / 0.1 $\textrm{m}{\ell}$/ guinea pig or 2.5 mg+CFA/0.1 $\textrm{m}{\ell}$/guinea pig) and topical application (2.5 mg/$\textrm{m}{\ell}$/guinea pig) with SDK or M291 at an interval of 1 week, and 2 weeks later, challenged by topical application with 25 mg/$\textrm{m}{\ell}$/guinea pig. SDK and M291 did not induce any reactions, showing 0 point of sensitization score and 0% of sensitization rate. In conclusion, it is suggested that SDK and M291 do not induce delayed contact hypersensitivity. In PCA test, rats were administered id with mouse anti-SDK serum and challenged iv with a mixture of antigen SDK and Evan's blue. SDK did not induce blue spots at the injection sites of both high (2.5 mg/mouse) and low (1.25 mg/mouse) dose-induced antisera. In contrast, BSA, positive control produced spots larger than 5 mm in diameter at the injection sites of BSA-induced antiserum up to $2^2$ ~ $2^4$dilution. In conclusion, it is suggested that SDK do not induce IgE production and is not a PCA-reaction inducer.

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Evaluation of a Rapid Immunochromatography Technique for Determination of Cd in Soils and Rice Grains (면역크로마토그래피를 이용한 토양 및 쌀의 Cd 간이진단법 평가)

  • Lee, Sang-Phil;Kim, Rog-Young;Abe, Kaoru;Kim, Sung-Chul;Kim, Won-Il;Yang, Jae E.
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.900-903
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    • 2012
  • Cadmium is known to be very toxic to human health and can be relative easily translocated from soils in plants. Therefore, a rapid method for screening Cd in soils and crops has become more and more important. For this reason, we examined a rapid immunochromatograpy (ICG) test kit which uses antigen-antibody reaction based on immunoassay and chromatography. Soils and rice grains collected from mine waste-contaminated sites were determined for their Cd contents using this kit. For comparison purposes, 0.1 M HCl and ICP-OES were employed as a conventional extraction and determination method. Cadmium contents in rice grains determined using ICG technique were $0.46{\sim}2.39mg\;kg^{-1}$ and Cd contents determined using 0.1 M HCl and ICP-OES were $0.52{\sim}1.97mg\;kg^{-1}$. The correlation between these two Cd contents were statistically significant ($r^2$=0.930). The results of Cd contents in soils also showed a statistically significant relationship between these two methods ($r^2$=0.975). On the basis of these results, ICG technique can be applied to rapidly quantify Cd in crops and soils. However, further research is necessary to apply ICG technique for the field screening.

Development and Clinical Evaluation of a Rapid Serodiagnostic Test for Toxoplasmosis of Cats Using Recombinant SAG1 Antigen

  • Chong, Chom-Kyu;Jeong, Woo-Seog;Kim, Hak-Yong;An, Dong-Jun;Jeoung, Hye-Young;Ryu, Jeong-Eun;Ko, A-Ra;Kim, Yong-Joo;Hong, Sung-Jong;Yang, Zhaoshou;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.207-212
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    • 2011
  • Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.

Clinical Application of Serum Tumor Abnormal Protein (TAP) in Colorectal Cancer Patients

  • Wu, Xue-Yan;Huang, Xin-En
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3425-3428
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    • 2015
  • Objective: To explore the association of serum tumor abnormal protein (TAP) with other serological biomarkers e.g. carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), carbohydrate antigen 19-9 (CA19-9) and its clinical application in colorectal cancer (CRC) patients. Methods: Patients (N=98) were enrolled into this study with histologically or cytologically confirmed CRC. Using a test kit, the level of TAP was determined, while chemiluminescence was used to measure the levels of some other common serological biomarkers e.g. CEA, CA125 and CA19-9. Results: The area of TAP condensed particulate matter decreased after chemotherapy compared with before chemotherapy when CT or MRI scans showed disease control. In contrast, it increased with disease progression (P<0.05). Furthermore, a statistically significant difference was confirmed in monitoring of TAP and common serological biomarkers e.g. CEA and CA19-9 (p<0.05). Conclusions: Detecting TAP in CRC patients has high sensitivity and specificity and can be used as a new independent indicator for clinically monitoring CRC patients in the course of chemotherapy.

Development of a Test Strip Reader for a Lateral Flow Membrane-based Immunochromatographic Assay

  • Park, Je-Kyun;Kim, Suhyeon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.127-131
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    • 2004
  • A low-cost, simple strip reader system using a linear movement mechanism of CD-ROM deck has been developed to characterize a lateral flow membrane-based immunochromatographic assay. The test strip reader was assembled by a CD-ROM deck and home-made optical head especially designed for immunoassays. The optical head for detecting reflected light from the test strip surface consists of green light-emitting diode, large area silicon photodiode, and anodized aluminum mounting block providing a slit structure for cutting light from the LED. The stepping motor of the deck was operated in the full step mode, whose distance of each reading point is about 0.15mm. The performance of the strip reader was tested by analysis of HBV(hepatitis B virus) antigen test kit. This strip reader can be useful for inexpensive, disposable, and membrane-based assays that provide visual evidence of the presence of an analyte in a liquid sample.

Prevalence of canine heartworm infection among dogs on breeding farms in Chungnam province (충남지역 집단 번식농장 사육견의 심장사상충 감염률 조사)

  • Lee, Jong-Hoon;Sim, Sang-Won;Kim, Hui;Kim, Young-Jin;An, Shin-Uk;Han, Kwei-Sup
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.19-26
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    • 2003
  • This study was attempted to survey on the prevalence of canine heartworm(Dirofilaria immitis) infection among 100 dogs(male 39, female 61) on the nine breeding farms in eastern Chungnam province in December 2002. Blood samples taken from dogs were examined for the presence of D immitis microfilaria by the modified Knott's test and an antigen test was using FASTest$\^$/ HW Antigne kit (Mega Cor A-6912 Horbranz-Austraia). 1. Eleven(11.0%) of the 100 examined dogs were microfilaria positive, while nineteen dogs(19.0%) were antigen positive, which suggested that the antigen test was more sensitive than the microfilarial test in detecting heartworm infection. 2. Infected dogs were observed higher more at 2 years older ages(4/48, 8.3%) and male(9/39, 23.1%) than young ages(4/48, 8.3%) and female(10/61, 16.4%). 3. The regional infection rates were of Gongju(15/43, 34.9%), Geumsan(4.27, 14.8%), while none of infection dogs in Yeongi(0/30, 0%). 4. Survey for hematological values of D immitis infected dogs : WBC and eosinophils were 21.4${\pm}$7.2 k/${\mu}\ell$, 3.5${\pm}$0.4 k/${\mu}\ell$, respectively. In conculsion, this study could be overemphasized the importance of control program the heartworm in eastern Chungnam province

Ebola Hemorrhagic Fever Outbreaks: Diagnosis for Effective Epidemic Disease Management and Control (에볼라 출혈열 발병 : 효과적인 전염병 관리 및 통제를 위한 진단)

  • Kang, Boram;Kim, Hyojin;Macoy, Donah Mary;Kim, Min Gab
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.87-92
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    • 2017
  • The first Ebola hemorrhagic fever outbreak occurred in the Democratic Republic of Congo and Sudan in 1976 and then emerged in West Africa in 2014 with a total of 27,741 cases and 11,284 deaths. The fever is caused by the Ebola virus, which belongs to the Filoviridae family and contains a ssRNA genome. The known subtypes of the virus are Bundibugyo ebolavirus, Reston ebolavirus, Sudan ebolavirus, $Ta\ddot{i}$ Forest ebolavirus, and Zaire ebolavirus. The Ebola outbreak was historically originated majorly from the East and Central African tropical belt. The current outbreaks in West Africa caused numerous deaths and spread fear in global society. In the absence of effective treatment strategies and any vaccine, accurate diagnosis is the most important contributing factor in the management and control of the epidemic disease. WHO (World Health Organization) has announced emergency guidance for the selection and use of Ebola in in vitro diagnostic assays. Numerous companies and research institutions have studied the various diagnosis methods and identified four WHO procurement approved as diagnosis kits: RealStar Ebolavirus Screen RT-PCR kit 1.0 (Altona), Liferiver-Ebola Virus (EBOV) Real time RT-PCR kit, Xpert Ebola Assay, and ReEBOV Antigen Rapid Test Kit. The efficiency of novel diagnostic kits such as Rapid Diagnosis Test (RDT) is currently being evaluated.

Comparative Analysis of the Multiple Test Methods for the Detection of Pandemic Influenza A/H1N1 2009 Virus

  • Choi, Young-Jin;Nam, Hae-Seon;Park, Joon-Soo;Kim, Hwi-Jun;Park, Kyung-Bae;Jeon, Min-Hyok;Kim, Chang-Jin;HwangBo, Young;Park, Kwi-Sung;Baek, Kyoung-Ah
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1450-1456
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    • 2010
  • Accurate and rapid diagnosis of Pandemic Influenza A/H1N1 2009 virus (H1N1 2009) infection is important for the prevention and control of influenza epidemics and the timely initiation of antiviral treatment. This study was conducted to evaluate the performance of several diagnostic tools for the detection of H1N1 2009. Flocked nasopharyngeal swabs were collected from 254 outpatients of suspected H1N1 2009 during October 2009. This study analyzed the performances of the RealTime Ready Inf A/H1N1 Detection Set (Roche), Influenza A (H1N1) Real-Time Detection Kit (Bionote), Seeplex Influenza A/B OneStep Typing Set [Seeplex Reverse Transcriptase PCR (RT-PCR)], BinaxNow Influenza A & B Test Kit [Binax Rapid Antigen Test (RAT)], and SD BIOLINE Influenza Ag kit (SD RAT). Roche and Bionote real-time RT-PCR showed identical results for the H1N1 2009 hemagglutinin gene. Compared with real-time RT-PCR, the sensitivities and specificities were 83.7% and 100% for Seeplex RT-PCR, 64.5% and 94.7% for Binax RAT, and 69.5% and 100% for SD RAT. The sensitivities of Seeplex RT-PCR, Binax RAT, and SD RAT in patients aged over 21 years were 73.7%, 47.4%, and 57.9%, respectively. The sensitivities of Seeplex RT-PCR, Binax RAT, and SD RAT on the day of initial symptoms were mostly lower (68.8%, 56.3%, and 31.3%, respectively). In conclusion, multiplex RT-PCR and RAT for the detection of H1N1 2009 were significantly less sensitive than real-time RT-PCR. Moreover, a negative RAT may require more sensitive confirmatory assays, because it cannot be ruled out from influenza infection.

A New IgG Immunoblot Kit for Diagnosis of Toxoplasmosis in Pregnant Women

  • Khammari, Imen;Saghrouni, Fatma;Lakhal, Sami;Bouratbine, Aida;Said, Moncef Ben;Boukadida, Jalel
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.493-499
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    • 2014
  • The determination of the accurate immune status of pregnant women is crucial in order to prevent congenital toxoplasmosis. Equivocal results with conventional serological techniques are not uncommon when IgG titers are close to the cut-off value of the test, so that a confirmatory technique is needed. For this purpose, we developed a homemade immunoblot (IB) using soluble extract of Toxoplasma gondii tachyzoites and assessed it by testing 154 positive, 100 negative, and 123 equivocal sera obtained from pregnant women. In order to select the more valuable bands in terms of sensitivity and specificity, we used the Youden Index (YI). The highest YIs were those given by the 32, 36, 98, 21, and 33 bands. The simultaneous presence on the same blot of at least 3 bands showed a much higher YI (0.964) and was adapted as the positivity criterion. The analysis of results showed that our homemade IB correlated well with the commercial LDBIO Toxo II $IgG^{(R)}$ kit recently recommended as a confirmatory test (96.7% of concordance).