• Title/Summary/Keyword: ELISA kit

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Evaluation of Two ELISA and Two Indirect Hemagglutination Tests for Serodiagnosis of Pulmonary Hydatid Disease

  • Eris, Fatma Nur;Akisu, Ciler;Aksoy, Umit
    • Parasites, Hosts and Diseases
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    • v.47 no.4
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    • pp.427-429
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    • 2009
  • To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.

Enzyme Immunoassay for the Sulfamethazine Residues in Pork Tissue

  • Park, Jun-Hong;Lim, Yoon-Kyu
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.287-290
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    • 1996
  • To control the maximum residue level (MRL) for sulfamethazine (SMZ) residues in pork tissue, a microbial inhibition method is a regulatory screening assay method in Korea. Microwell plate-based competitive enzyme immunoassay (ELISA) kit is avalable for routine screening of SMZ residues in pork tissue. One ELISA kit is evaluated. Phosphate buffer extracts of samples fortified with SMZ at 0, 1, 5, and 10 ng/g were used in a recovery test of the kit. Market pork samples were assayed by the kit. Recovery of sulfamethazine was 104% at 10 ng/g. Intraassay variations and interassay variations for the kit were 7.70% and 5.76%, respectively. Concentration causing 50% inhibition of color development compared with blanks was 16.4ng. The violative pork samples with over MRL (0.1 $\mu\textrm{g}$/g) was 4 of 32 cases (12.5%) by used ELISA kit. This result indicates a possibility of the ELISA kit for screening test of SMZ residues in pork tissue, and still needs a comfirmatory assay for mandatory purposes.

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A comparative study on the diagnosis of ELISA test and PPD test of the bovine tuberculosis (ELISA를 이용한 우결핵검사 결과에 대한 PPD 접종법 결과 분석 비교)

  • Lee, Jong-Jin;Kim, Duk-Soon;Lee, Jong-Hwa;Lee, Cheong-San
    • Korean Journal of Veterinary Service
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    • v.33 no.4
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    • pp.335-340
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    • 2010
  • On the basis of the 2009 business plan, 20,394 Korean native cattle and beef cattle were carried examination of bovine tuberculosis by using ELISA technique from March to December. As a result, 66 cattle tested positive for tuberculosis and showed 0.32% positive ratio. Intradermal tuberculin test about 66 cases of ELISA positive cattle was carried out, and all of 66 cattle were confirmed as negative. However, when 7 PPD-positive cattle derived from slaughterhouse were tested by 20k ELISA kit and MS ELISA kit, 3 (2 suspect) cattle and 5 cattle showed positive results, respectively. As compared to the results of PPD test, the concordance rates were 43% (71% included suspect) with 20k ELISA kit and 71% with MS ELISA kit.

Studies on use of milk progesterone EIA-kit for diagnosis of reproductive disorders and non-pregnancy in dairy cows (젖소의 번식장애 및 비임신진단을 위한 Milk Progesterone 측정 EIA-kit의 이용에 관한 연구)

  • Chung, Byung-hyun;Lee, Byeong-han;Kang, Young-sun;Kim, Jin-young;Nam, Hyoung-young;Lee, Kang-yeol;Hwang, Yoon-sik;Yang, Kwang-hun;Chung, Kil-saeng
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.169-177
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    • 1995
  • This study was carried out to investigate the effect, range of practice, and propriety for diagnosis of early non-pregnancies and reproductive disorders by dairy cows' milk progesterone analysis used EIA-kit of home products. The results were summarized as follows : 1. During 2 to 6 months after artificial insemination, the results of milk progesterone measurement by Home-kit and Auto ELISA reader-kit with pregnant dairy cows(152 heads) certified by rectal palpation were revealed, in Home-kit, 145 heads(95.4%) of positive reaction, 7 heads(4.6%) of quasi-positive, and 0 heads(0%) of negative among 152 heads and, in Auto ELISA reader-kit, 152 heads(100%) of positive reaction among 152 heads. 2. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Home-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 147 heads(82.1%) among 179 heads of positive reaction by Home-kit and 5 heads(31.3%) among 16 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 42 heads(100%) among 42 heads of negative were non-pregnant. 3. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Auto ELISA reader-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 146 heads(86.9%) among 168 heads of positive reaction by Auto ELISA reader-kit and 6 heads(28.6%) among 21 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 48 heads (100%) among 48 heads of negative were non-pregnant. 4. For the accuracy of the rectal palpation, Home-kit and Auto ELISA reader-kit were used in the cows of ovarian diseases. The results were following : in the cows of reproductive disorders expected negative milk progesterone, the accuracies of rectal palpation were the same 75.5%(40 heads among 53 heads) by Home-kit and Auto ELISA reader-kit, and in the cows of reproductive disorders expected positive milk progesterone, the accuracies of rectal palpation were 82.6%(19 heads among 23 heads) and 91.3%(21 heads among 23 heads) by Home-kit and Auto ELISA reader-kit, respectively, and the general accuracies of rectal palpation were 77.6%(59 heads among 76 heads) and 80.3%(61 heads among 76 heads) by Home-kit and Auto ELISA reader-kit, respectively.

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Validation of PCR and ELISA Test Kits for Identification of Domestic Animal Species in Raw Meat and Meat Products in Korea (국내 유통 식육 및 식육가공품에서 축종감별을 위한 PCR 및 ELISA 검사법 검증)

  • Heo, Eun-Jeong;Ko, Eun-Kyung;Seo, Kun-Ho;Kim, Young-Jo;Park, Hyun-Jung;Wee, Sung-Hwan;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.29 no.2
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    • pp.158-163
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    • 2014
  • In this study, two commercial PCR and ELISA test kits were examined for identification of eight animal species (beef, pork, chicken, duck, turkey, goat, lamb, and horse) from raw meat and meat products in Korea. The detection limit in RAW meat ELISA kit$^{(R)}$ on three types of meat samples blended with beef, pork and chicken, demonstrated that all meat species were differentiable down to 0.2%. RAW meat ELISA kit$^{(R)}$ on animal species resulted in differentiation rate of 94.5% for beef, 93.3% for pork, 90% for lamb, and 100% for chicken, duck, turkey, goat, and horse. In contrast, Powercheck Animal Species ID PCR kit$^{TM}$ resulted in 100% specificity at 0.05% limit of detection for all meat species. The detection limit of Cooked Meat ELISA kit$^{(R)}$ on mixed meat samples heat-treated with different temperatures and times, resulted in 0.1% for all heat-treated mixed meat except for chicken at 1.0%. Additionally, ELISA kit on sixty meat products resulted in specificity of 31.8% for ham, 13.6% for sausages, and 12.5% for ground processed products, and relatively low rate for more than 2 types of mixed meats. On the contrary, meat species differentiation using PCR kit showed higher percentage than that using ELISA kit$^{(R)}$: 50.0% for ham, 41.7% for sausages, and 28.6% for ground processed meat. Futhermore, PCR kit on 54 dried beef meats detected pork genes in 13 products whereas ELISA kit showed negative results for all products. Hence, the possibility of cross-contamination during manufacturing process was investigated, and it was found that identical tumblers, straining trays, cutters and dryers were used in both beef and pork jerky production line, suggesting the inclusion of pork genes in beef products due to cross-contamination. In this study, PCR and ELISA test kits were found to be excellent methods for meat species differentiation in raw meat and heat-processed mixed meat. However, lower differentiation rate demonstrated in case of meat processed products raised the possibility of inclusion of other species due to cross-contamination during manufacturing process.

Serodiagnosis of Extraintestinal Amebiasis: Retrospective Evaluation of the Diagnostic Performance of the Bordier® ELISA Kit

  • Beyls, Nicolas;Cognet, Odile;Stahl, Jean-Paul;Rogeaux, Olivier;Pelloux, Herve
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.71-74
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    • 2018
  • Soluble antigens from an axenic culture of Entamoeba histolytica were used to develop a commercial ELISA kit to quantify anti-E. histolytica antibodies in sera of patients with extraintestinal amebiasis in non-endemic settings. The diagnostic specificity and sensitivity of the test were assessed retrospectively using 131 human serum samples with amoebic serologic status available. They were selected according to their results in immunofluorescence (IFAT) and were separated in 2 sample categories: 64 sera with positive results by IFAT and 67 with negative results by IFAT. The sensitivity and specificity of the ELISA kit were assessed at 95.0% and 94.0% compared to the IFAT. The test can be useful to exclude a potential diagnosis of amebiasis and could be used as a screening method since ELISA is an automated technique.

Evaluation of an ELISA kit for the Serodiagnosis of Pulmonary Tuberculosis by Using Mixed Antigens of Mycobacterium Tuberculosis (폐결핵진단에서 결핵균 혼합항원을 이용한 혈청학적 검사의 유용성에 관한연구)

  • Park, Seung-Kyu;Kim, Phil-Ho;Kim, Seung-Chul;Choi, In-Hwan;Cho, Sang-Nae;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.558-567
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    • 2000
  • Background : Recently, serologic techniques for tuberculosis have been developed and some of them, which are focusing on detection of serum antibodies mainly directed against specific 38-kDa Mycobacterium tuberculosis, have already been introduced into the markel. In this study, diagnostic significance of a new serologic test(ELISA kit) for pulmonary tuberculosis was evaluated. Method : Serologic test with newly developed ELISA kit was performed upon 474 individuals, who include 333 active pulmonary tuberculosis patients, 80 healthy cases, and 61 tuberculosis contact cases. This serologic test was based on the ELISA technique and designed to detect antibodies to mixed complex antigens including 38-kDa, which were developed by Erume Biotech Co., Seoul. Active pulmonary tuberculosis was diagnosed by sputum AFB smear and culture methods. Results : The seropositivities using this ELISA kit were 82.1% and 73.6% in smear-positive and negative groups among active pulmonary tuberculosis, respectively. And, it also showed that seronegativities were 97.5% and 85.2% in healthy and contact groups, respectively. As a whole, the results of our study using the ELISA kit as a diagnostic method for pulmonary tuberculosis showed 80.0% sensitivity for active pulmonary tuberculosis, 97.5% specificity, 96.1% positive predictive value, and 65.0% negative predictive value when the prevalence of tuberuclosis in the samples was 60.1%. Conclusion : Our results reveal that the detection of antibody its reaction with 38-kDa antigen of M. tuberculosis is not sufficient to be accepted as single diagnostic method for pulmonary tuberculosis. However, they suggest that ELISA kit may be considered as an adjunctive test to standard diagnostic techniques of pulmonary tuberculosis.

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Correlation Between Skin Prick Test and Enzyme-linked Immunosorbent Assay Using Serum for Identification of Subjects Positive to Major Indoor Respiratory Allergens (실내환경 주요 호흡기 알레르기 항원 감작검사를 위한 피부단자 시험과 혈청 ELISA시험의 상관성에 대한 연구)

  • Heo, Yong;Kim, Hyoung-Ah
    • Journal of Environmental Health Sciences
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    • v.34 no.5
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    • pp.369-373
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    • 2008
  • The indoor environment is an important source of exposure to various aeroallergens and pathogenic microorganism. It has been shown that exposure to aeroallergens enhances the risk of indoor inhabitants developing asthma. Since the skin prick test, a typical clinical method for identification of subjects positive to allergens, can rarely cause fatal or non-fatal reactions in susceptible persons, an in vitro assay such as ELISA using serum has been considered for testing positivity against various allergens. We evaluated the validity of a serum ELISA kit for screening positive subjects to major aeroallergens including Dermatophagoides farinae, Dermatophagoides pteronyssinus, cockroach, Alternaria, Cladosporium, Aspergillus, Penicillium, dog hair, cat fur, mugwort, and ragweed. The ELISA results were compared with the skin prick test results, and sensitivity, specificity, and overall accuracy were calculated to each allergen. Higher sensitivities were obtained from D. farinae, (77.8%) and D. pteronyssinus (69.2%), but sensitivities to Aspergillus, Penicillium, dog hair, cat fur, and ragweed were very low down to 0%. Specificity ranged from 88.7% (cat fur) to 100% (mugwort and ragweed). Overall the accuracy of the serum ELISA kit was relatively high, in that the lowest was 85.1% for cat fur and the highest was 98.6% for Alternaria, Cladosporium, and ragweed. Considering specificity and overall accuracy for the serum ELISA kit, it may be considered reliable. However, when the kit is used for screening purpose, positivity to aeroallergens should be carefully determined since sensitivity for the kit is low.

Development of ELISA for cortisol and it's application to clinical use (ELISA를 이용한 cortisol 측정법의 정립 및 임상적 응용)

  • Na, Ki-jeong;Lee, Chang-woo
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.731-741
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    • 1996
  • ELISA kit for cortisol was developed and then evaluated. Polyclonal antihydrocortisone-3-(o-carboxymethyl)oxime BSA rabbit serum was used to coat the 96-well microplates. The minimum detection limit of the kit was 250pg of cortisol per milliliter. The within-run variation and the day to day variation of the ELISA system were 2.0 and 5.9 at maximum, respectively. The kit was used to determine whether salivary cortisol concentration could replace blood cortisol concentration in dexamathasone suppression test of dogs. Changes of cortisol concentration were measured in serum or saliva after intravenous administration of 0.01mg of dexamethasone per kilogram of body weight. Blood alone, saliva alone or both were collected at 0, 30, 60, 120, 240, and 360 minutes after injection of dexamethasone. The change in blood cortisol concentration was found to be suitable in dexamathasone suppression test of dogs, but the change in salivary cortisol concentration was not. The kit was also used to determine whether salivary cortisol concentration could be a stress index as well as blood cortisol concentration in dogs. Two types of trial were performed to estimate the stress either by blood or salivary cortisol concentration. The first trial was stress experiment by intravenous injection of 0.2IU of PZI-insulin per kilogram body weight. Either blood alone or saliva alone was collected at 0, 30, 60, and 90 minutes after insulin administration. Both blood and salivary cortisol concentration were found to be suitable index in estimating stress from hypoglycemia by injection of insulin. The second trial was stress experiment by electrical irritation. The dogs were irritated with anti-bark device for 10 seconds. Blood was collected before and at 2 and 5 minutes after electrical irritation. Saliva was collected before and at 3 and 6 minutes after electrical irritation. The blood cortisol concentration, but not the salivary cortisol concentration was found to be suitable index in estimating stress from electrical irritation. Cushing syndrome in a dog was also successfully diagnosed with this kit.

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Development of an ELISA kit for the detection of residual sulfadimethoxine in edible animal products (축산물 잔류 sulfadimethoxine 검출용 ELISA kit 개발)

  • Kim, Woo-taek;Kim, Seong-hee;Yoon, Byoung-su;Lim, Yoon-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.601-609
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    • 2000
  • An enzyme linked immunosorbent assay (ELISA) was developed to screen residues of sulfadimethoxine (SDM) in edible animal products. An indirect competitive ELISA was allowed to compete with rabbit anti-SDM for binding to a limited amount of SDM-gelatin conjugate and SDM in serum samples. Sera was diluted 20 times with phosphate buffered saline (PBS) and boiled for 5 minutes to destruct immunoglobulins of serum. Detection limit of this competitive ELISA for SDM was 0.1 ppb or less. Among eight sulfonamide analogues tested for specifity, only sulfamonomethoxine showed significant cross-reaction in the assay. The EC-50 value for sulfamonomethoxine was 3.5 ppm. Recovery of SDM in spiked serum samples between 100 ppb and 500 ppb ranged from 110.7% to 128.9%.

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