Displaying publications 1 - 20 of 495 in total

  1. Lakshmipriya T, Gopinath SC, Tang TH
    PLoS ONE, 2016;11(3):e0151153.
    PMID: 26954237 DOI: 10.1371/journal.pone.0151153
    Enzyme Linked Immunosorbent Assay (ELISA) is the gold standard assay for detecting and identifying biomolecules using antibodies as the probe. Improving ELISA is crucial for detecting disease-causing agents and facilitating diagnosis at the early stages of disease. Biotinylated antibody and streptavidin-conjugated horse radish peroxide (streptavidin-HRP) often are used with ELISA to enhance the detection of various kinds of targets. In the present study, we used a competition-based strategy in which we pre-mixed free biotin with streptavidin-HRP to generate high-performance system, as free biotin occupies some of the biotin binding sites on streptavidin, thereby providing more chances for streptavidin-HRP to bind with biotinylated antibody. ESAT-6, which is a protein secreted early during tuberculosis infection, was used as the model target. We found that 8 fM of free biotin mixed with streptavidin-HRP anchored the higher detection level of ESAT-6 by four-fold compared with detection without free biotin (only streptavidin-HRP), and the limit of detection of the new method was 250 pM. These results suggest that biotin-streptavidin competition can be used to improve the diagnosis of analytes in other types of sensors.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Enzyme-Linked Immunosorbent Assay/standards
  2. Anuar H, Greer GJ, Ow-Yang CK, Sukumaran KD
    PMID: 6398916
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay*
    Sains Malaysiana, 2012;41:423-430.
    Sistem minigenom telah digunakan untuk mengkaji replikasi dan transkripsi virus RNA tidak bersegmen. Objektif kajian ini adalah untuk membina sistem minigenom bagi virus NDV strain tempatan, AF2240 serta bagi mengkaji mekanisme transkripsi dan replikasi virus ini. Bagi tujuan ini lima plasmid digunakan iaitu pMGNDV, pCITENP, pCITEP, pTriEX-T7, dan pGEML. Kesemua plasmid diekstrak secara berskala besar dan dimendakkan menggunakan polietilina glikol. Hasil ekstrak ini digunakan untuk transfeksi ke dalam sel. Translasi in vitro dilakukan dengan menggunakan pCITENP, pCITEP, dan pTriEX-T7 untuk memastikan kesemua konstruk ini berfungsi. Hasil pemblotan western menunjukkan protein bersaiz ~100 kDa (T7), ~53 kDa (NP), ~53 dan 55 kDa (P) berjaya diekspreskan. Protein CAT diperoleh apabila plasmid yang mengekodkan minigenom NDV ditransfeksi bersama plasmid yang mengekodkan protein nukleokapsid (NP), fosfoprotein (P) dan subunit besar polimerase (L) ke dalam sel BHK-21. Dianggarkan 55 pg protein CAT berjaya diperoleh menggunakan kit CAT ELISA. Hasil pemblotan western turut menunjukkan protein CAT bersaiz 25 kDa dihasilkan. Kesimpulannnya, system minigenom ini berupaya untuk berfungsi dan mampu mengekspreskan gen asing di dalam sel mamalia BHK-21.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  4. Sue MJ, Yeap SK, Omar AR, Tan SW
    Biomed Res Int, 2014;2014:653014.
    PMID: 24971343 DOI: 10.1155/2014/653014
    Polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) is an immunodetection method that can quantify PCR product directly after immobilization of biotinylated DNA on a microplate. This method, which detects nucleic acid instead of protein, is a much more sensitive method compared to conventional PCR method, with shorter analytical time and lower detection limit. Its high specificity and sensitivity, together with its semiquantitative ability, give it a huge potential to serve as a powerful detection tool in various industries such as medical, veterinary, and agricultural industries. With the recent advances in PCR-ELISA, it is envisaged that the assay is more widely recognized for its fast and sensitive detection limit which could improve overall diagnostic time and quality.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  5. Nazaimoon W, Ng ML, Bak K
    Malays J Pathol, 1993 Jun;15(1):75-83.
    PMID: 8277795
    A simple, non-isotopic in-house enzyme-linked immunoabsorbant assay (ELISA) for human growth hormone (GH) was developed. The assay involved using in-house polyclonal anti-GH adsorbed onto 96-well microtitre plates, commercially prepared mouse monoclonal anti-GH, and goat anti-mouse IgG horseradish peroxidase detection system. Results of recovery and parallelism studies ranged from 95%-106% and 98%-101% respectively, of the expected values. The detection limit of the assay was 0.008 mIU/well or the equivalent to 0.4 mIU/L of undiluted serum. Intra- and interassay coefficients of variations were 4.8%-7.9% and 6.5%-8.7% respectively. Serum GH levels measured in this assay correlated well with those measured in established in-house radioimmunoassays (r = 0.985, p < 0.001) and immunoradiometric assay from NETRIA (r = 0.984, p < 0.001).
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  6. Goh KH, Goh ML, Thean ET, Khalid BA
    Med. J. Malaysia, 1992 Dec;47(4):248-60.
    PMID: 1303476
    A supersensitive ELISA was developed for measurement of thyroid-stimulating hormone (TSH) concentrations in serum using in-house rabbit polyclonal antisera and a commercial monoclonal antibody. The assay was optimised and validated by recovery, linearity and cross-reactivity experiments and further compared to other available assays and EQAS samples. Good precision was obtained with a working assay range of 0.2 to 100 mIU/L with < 10% coefficient of variation (CV) for both intra and interassay. The assay is highly sensitive and specific with a minimum detectable limit of 0.07 mIU/L and negligible cross-reactivities against LH, FSH, HCG and other pituitary peptides. Good correlations were obtained when compared to Abbott hTSH EIA (r = 0.993; p < 0.001; n = 85) and NETRIA IRMA (r + 0.995; p < 0.001; n = 76). The normal reference range established was 0.4 to 4.0 mIU/L (n = 76). TSH levels in serum of thyrotoxic patients (n = 83) were significantly lower (0.07 to 0.20 mIU/L, p < 0.0001) and completely distinct from normal values thereby obviating the requirement of a TRH-stimulation test. Stability studies showed that coated wells can be stored at 4 degrees C for at least 2 months. This highly sensitive in-house hTSH ELISA which is cheap, stable and readily available is useful for diagnosis and management of patients with various thyroid disorders.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  7. Cheng HM, Ngeow YF, Sam CK
    J. Immunol. Methods, 1989 Nov 30;124(2):235-8.
    PMID: 2600427
    Heat treatment of sera at 56 degrees C for 30 min results in positive ELISA reactions for anti-cardiolipin antibody (aCL) in sera that had undetectable or low levels of aCL before heat inactivation. The positive, potentiated reactivity of the heated sera in the aCL ELISA could be inhibited with the cardiolipin antigen and was abolished by prior IgG depletion using staphylococcal protein A. The heat-potentiating effect of aCL binding in ELISA was evident in both normal human sera and clinical sera including sera from patients with systemic lupus erythematosus and syphilis.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay*
  8. Lakshmipriya T, Gopinath SCB, Hashim U, Murugaiyah V
    Int. J. Biol. Macromol., 2017 Dec;105(Pt 1):796-800.
    PMID: 28732727 DOI: 10.1016/j.ijbiomac.2017.07.115
    Enzyme Linked Immunosorbent Assay (ELISA) is a standard assay that has been used widely to validate the presence of analyte in the solution. With the advancement of ELISA, different strategies have shown and became a suitable immunoassay for a wide range of analytes. Herein, we attempted to provide additional evidence with ELISA, to show its suitability for multi-analyte detection. To demonstrate, three clinically relevant targets have been chosen, which include 16kDa protein from Mycobacterium tuberculosis, human blood clotting Factor IXa and a tumour marker Squamous Cell Carcinoma antigen. Indeed, we adapted the routine steps from the conventional ELISA to validate the occurrence of analytes both in homogeneous and heterogeneous solutions. With the homogeneous and heterogeneous solutions, we could attain the sensitivity of 2, 8 and 1nM for the targets 16kDa protein, FIXa and SSC antigen, respectively. Further, the specific multi-analyte validations were evidenced with the similar sensitivities in the presence of human serum. ELISA assay in this study has proven its applicability for the genuine multiple target validation in the heterogeneous solution, can be followed for other target validations.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
  9. AbuHassan KJ, Bakhori NM, Kusnin N, Azmi UZM, Tania MH, Evans BA, et al.
    Conf Proc IEEE Eng Med Biol Soc, 2017 07;2017:4512-4515.
    PMID: 29060900 DOI: 10.1109/EMBC.2017.8037859
    Tuberculosis (TB) remains one of the most devastating infectious diseases and its treatment efficiency is majorly influenced by the stage at which infection with the TB bacterium is diagnosed. The available methods for TB diagnosis are either time consuming, costly or not efficient. This study employs a signal generation mechanism for biosensing, known as Plasmonic ELISA, and computational intelligence to facilitate automatic diagnosis of TB. Plasmonic ELISA enables the detection of a few molecules of analyte by the incorporation of smart nanomaterials for better sensitivity of the developed detection system. The computational system uses k-means clustering and thresholding for image segmentation. This paper presents the results of the classification performance of the Plasmonic ELISA imaging data by using various types of classifiers. The five-fold cross-validation results show high accuracy rate (>97%) in classifying TB images using the entire data set. Future work will focus on developing an intelligent mobile-enabled expert system to diagnose TB in real-time. The intelligent system will be clinically validated and tested in collaboration with healthcare providers in Malaysia.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  10. Syed-Hussain SS, Howe L, Pomroy WE, West DM, Smith SL, Williamson NB
    Vet. Parasitol., 2014 Jun 16;203(1-2):21-8.
    PMID: 24582279 DOI: 10.1016/j.vetpar.2014.01.003
    Recent reports indicate Neospora caninum has a possible role in causing abortions in sheep in New Zealand. Knowledge about the epidemiology of neosporosis in sheep is limited. This study aimed to adapt and validate a commercially available ELISA assay as an IgG avidity assay to discriminate between acute (primary and re-inoculated) and chronic N. caninum infections in sheep. In addition, it was used to compare the antibody avidity values between lambs from ewes inoculated with N. caninum either during the pregnancy or in the previous year. The avidity assay was undertaken by using 6M urea for the first wash after incubation with the primary antibody in the commercial ELISA (Chekit* Neospora antibody test kit, IDEXX Laboratories, Australia). Sequential serum samples were obtained from naïve ewes (n=16) experimentally inoculated with live N. caninum tachyzoites. All ewes were seropositive by two weeks post-inoculation and remained seropositive for 20 weeks post-inoculation. There was a linear relationship between time after inoculation and avidity values (p<0.05) over the first 24 weeks. In Week 4, all animals had avidity values <35% and by Week 8, 8/16 animals had avidity values of >35%. These results suggest that an avidity value of <35% indicates a recent primary infection while a value of >35% is indicative of a chronic infection. The assay was then validated using samples from other groups of experimentally inoculated sheep as well as samples from naturally infected ewes. When comparing sample to positive ratio (S/P) and avidity values from lambs born from recently inoculated ewes with those from ewes inoculated the previous year and re-inoculated in the current year, it was possible to differentiate the lambs at 2 weeks of age. Lambs from recently inoculated ewes had low S/P and avidity values at 2 weeks of age which increased by 12 weeks of age. In comparison, lambs from re-inoculated ewes had high S/P and avidity values at 2 weeks of age, due to maternal antibody influence but values were similar to those from lambs that were born from recently inoculated ewes at 12 weeks of age. Avidity values for four naturally infected ewes were all >60% indicating chronic infection. These results suggest that the assay is able to discriminate between recent and chronic infection in sheep as well as able to differentiate lambs with maternal immunity compared to their own de novo immunity. As such it can be utilized to understand the kinetics of N. caninum infection in sheep.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/standards; Enzyme-Linked Immunosorbent Assay/veterinary*
  11. Teoh BT, Sam SS, Tan KK, Johari J, Abd-Jamil J, Hooi PS, et al.
    Sci Rep, 2016 06 09;6:27663.
    PMID: 27278716 DOI: 10.1038/srep27663
    Timely and accurate dengue diagnosis is important for differential diagnosis and immediate implementation of appropriate disease control measures. In this study, we compared the usefulness and applicability of NS1 RDT (NS1 Ag Strip) and qRT-PCR tests in complementing the IgM ELISA for dengue diagnosis on single serum specimen (n = 375). The NS1 Ag Strip and qRT-PCR showed a fair concordance (κ = 0.207, p = 0.001). While the NS1 Ag Strip showed higher positivity than qRT-PCR for acute (97.8% vs. 84.8%) and post-acute samples (94.8% vs. 71.8%) of primary infection, qRT-PCR showed higher positivity for acute (58.1% vs. 48.4%) and post-acute (50.0% vs.41.4%) samples in secondary infection. IgM ELISA showed higher positivity in samples from secondary dengue (74.2-94.8%) than in those from primary dengue (21.7-64.1%). More primary dengue samples showed positive with combined NS1 Ag Strip/IgM ELISA (99.0% vs. 92.8%) whereas more secondary samples showed positive with combined qRT-PCR/IgM ELISA (99.4% vs. 96.2%). Combined NS1 Ag Strip/IgM ELISA is a suitable combination tests for timely and accurate dengue diagnosis on single serum specimen. If complemented with qRT-PCR, combined NS1 Ag Strip/IgM ELISA would improve detection of secondary dengue samples.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods; Enzyme-Linked Immunosorbent Assay/standards
  12. Cheong KB, Cheong SK, Boo NY, Jemilah M, Ton SH
    Malays J Pathol, 1995 Dec;17(2):91-6.
    PMID: 8935133
    An in-house enzyme-linked immunoabsorbant assay (ELISA) for SP-A was successfully developed using in-house polyclonal anti SP-A and a commercial polyclonal anti-rabbit immunoglobulin horseradish peroxidase conjugate system. The standard curve, generated by using 50 ng of SP-A to coat the plate and 1:500 dilution of polyclonal anti SP-A as a primary antibody, was linear for concentrations of SP-A ranging from 4 micrograms/l to 4000 micrograms/l and reproducible. Results of recovery study of SP-A from a known sample of tracheal aspirate ranged from 94%-114%. Intra- and inter-assay coefficients of variations were 2.7% and 5.6% respectively for a known sample of tracheal aspirate. Interference study showed that tracheal aspirate did not interfere with the assay. The assay developed was intended to be used for SP-A measurement in tracheal aspirates obtained from neonates with and without respiratory distress syndrome.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Enzyme-Linked Immunosorbent Assay/standards
  13. Rahmah N, Lim BH, Khairul Anuar A, Shenoy RK, Kumaraswami V, Lokman Hakim S, et al.
    Trans. R. Soc. Trop. Med. Hyg., 2001 8 9;95(3):280-4.
    PMID: 11490997
    An IgG4 ELISA based on a novel recombinant antigen was evaluated for detection of Brugia malayi infection, using 2487 sera from various institutions: 2031 samples from Universiti Sains Malaysia, 276 blinded sera from 2 other institutions in Malaysia, 140 blinded sera from India and 40 blinded sera from Thailand. These sera were from various groups of individuals, i.e., microfilaraemics, chronic patients, endemic normals, non-endemic normals and individuals with other parasitic and bacterial infections. Based on a cut-off optical density reading of 0.300, the IgG4 ELISA demonstrated specificity rates of 95.6-100%, sensitivity rates of 96-100%, positive predictive values of 75-100% and negative predictive values of 98.9-100%. These evaluation studies demonstrated the high specificity and sensitivity of this test for the detection of active B. malayi infection. Thus, the IgG4 ELISA would be very useful as a tool in diagnosis and in elimination programmes for brugian filariasis.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Enzyme-Linked Immunosorbent Assay/standards
  14. Kapitonova MY, Kuznetsov SL, Salim N, Othman S, Kamauzaman TM, Ali AM, et al.
    Bull. Exp. Biol. Med., 2014 Jan;156(3):393-8.
    PMID: 24771384 DOI: 10.1007/s10517-014-2357-8
    Morphological and phenotypical signs of cultured readaptation osteoblasts were studied after a short-term space mission. The ultrastructure and phenotype of human osteoblasts after Soyuz TMA-11 space flight (2007) were evaluated by scanning electron microscopy, laser confocal microscopy, and ELISA. The morphofunctional changes in cell cultures persisted after 12 passages. Osteoblasts retained the drastic changes in their shape and size, contour deformation, disorganization of the microtubular network, redistribution of organelles and specialized structures of the plasmalemma in comparison with the ground control cells. On the other hand, the expression of osteoprotegerin and osteocalcin (bone metabolism markers) increased; the expression of bone resorption markers ICAM-1 and IL-6 also increased, while the expression of VCAM-1 decreased. Hence, space flight led to the development of persistent shifts in cultured osteoblasts indicating injuries to the cytoskeleton and the phenotype changes, indicating modulation of bone metabolism biomarkers.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  15. Raja Nhari RM, Hamid M, Rasli NM, Omar AR, El Sheikha AF, Mustafa S
    J. Sci. Food Agric., 2016 May;96(7):2524-31.
    PMID: 26611757 DOI: 10.1002/jsfa.7547
    Porcine blood is potentially being utilized in food as a binder, gelling agent, emulsifier or colorant. However, for certain communities, the usage of animal blood in food is strictly prohibited owing to religious concerns and health reasons. This study reports the development of monoclonal antibodies (MAbs) against heat-treated soluble proteins (HSPs) of autoclaved porcine blood; characterization of MAbs against blood, non-blood and plasma from different animal species using qualitative indirect non-competitive enzyme-linked immunosorbent assay (ELISA); and immunoblotting of antigenic components in HSPs of porcine blood.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  16. Ming CH, Fan YS
    J. Immunol. Methods, 1988 May 09;109(2):253-5.
    PMID: 3361136
    Addition of the non-ionic detergent Tween 20 to the serum diluent enhances anti-cardiolipin binding reactivity in an ELISA system. Maximal enhancement was obtained using a concentration of 0.05% Tween 20 in the diluent. Non-specific interactions were also considerably reduced.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  17. Ambrose JH, Sekaran SD, Azizan A
    J Trop Med, 2017;2017:8072491.
    PMID: 28740517 DOI: 10.1155/2017/8072491
    BACKGROUND: The proper management of patients infected with dengue virus requires early detection. Here, real-time molecular assays have proven useful but have limitations, whereas ELISAs that detect antibodies are still favored but results are obtained too late to be of clinical value. The production of DENV NS1 peaks early during infection and its detection can combine the advantages of both diagnostic approaches.

    METHODS: This study compared assays currently used for detecting DENV infection at the Florida Department of Health including anti-DENV IgM and IgG ELISAs as well as qRT-PCR, against a commercially available DENV NS1 ELISA. These comparisons were made among a group of 21 human sera.

    RESULTS: Nine of 14 (64.3%) DENV qRT-PCR+ samples were also DENV NS1+. Interestingly, the 5 NS1- samples that were qRT-PCR+ were additionally IgM- and IgG+ suggesting a nonprimary infection. Compared to qRT-PCR, the NS1 assay had a sensitivity of 64.3%, specificity 100%, PPV of 100%, and NPV of 58.3%.

    CONCLUSIONS: The NS1 ELISA performed as expected in known DENV qRT-PCR+ samples; however negative NS1 results for qRT-PCR+ and IgG+ sera seemingly reduced the usefulness of the NS1 ELISA for nonprimary cases. We therefore conclude that diagnosis obtained via DENV NS1 ELISA deserves further investigation.

    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  18. Shamsul, B.S., How Pai, S.
    Homocysteine could be a mechanism that underlies the effects of lead on cardiovascular system. This study aims to identify the relationship between lead exposure and homocysteine levels among workers. A comparative cross-sectional study was carried out on 80 workers of an automotive components manufacturing factory; that comprised of 40 exposed workers and 40 non-exposed workers. Blood samples of respondents were taken by fingerprick. The blood samples were analyzed for blood lead concentration by using Atomic Absorption Spectrometry Graphite Furnace Model GBC 908AA. Besides that, ELISA Kit was used to show the homocysteine level among the respondents. Questionnaires were used to obtain demography information of respondents. Results from the statistical analysis showed that the mean blood lead concentration for exposed respondents was 5.53±4.74 μg/dL and 3.53±2.81 μg/dL for the comparative respondents. Mann-Whitney U test showed that there was no significance difference between the mean blood lead concentration of the exposed and comparative group (z=-1.178; p=0.075). The blood lead concentration ranged 0.68-17.95 among the exposed group and with a range of 0.084-11.96 for the comparative group. The mean homocysteine level (μmol/L) was 32.48±2.481μmol/L for the exposed group and 16.50±4.0960 μmol/L for the comparative group. There was a significant difference in homocysteine level (μmol/L) between the exposed (32.48±2.481) and comparative (16.50±4.0959) groups (z = -7.699, p
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  19. Shamsul, B.S., Zakirah, M.
    The main objective of this study is to determine the association between respirable hexavalent chromium compounds with urinary β2-microglobulin levels among welders in an automotive components manufacturing plant. 49 welders and 39 workers involved in stamping process were selected as the exposed and the comparative group. β2-microglobulin is a protein renal tubular dysfunction marker that can indicate renal dysfunction caused by heavy metal. Air samples of worker’s breathing zone were collected using personal air sampling pump and filter papers. Filter papers were then diluted and analysed with Atomic Absorption Spectrophotometry (AAS). Workers’ urine samples were collected at the end of 8-hour work shift and analysed with β2-microglobulin ELISA Kit (IBL-Hamburg) and a microtiter reader. Meanwhile, creatinine levels were analysed with creatinine test strips and Reflotron®. A mean concentration of respirable hexavalent chromium compounds in air for the exposed group was 0.135 ± 0.043μg/m3 while for the non-exposed group was 0.124 ± 0.029μg/m3. The mean level of urinary β2-microglobulin per creatinine for the exposed group was 84.996 ± 39.246μg/g while that of the comparative group was 61.365 ± 21.609μg/g. The concentrations of respirable hexavalent chromium compounds were higher in the exposed group compared to the comparative group (Z=-2.444, p=0.015). β2-microglobulin level was also higher in the exposed group compared to the non-exposed group (t=3.821, p=
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
  20. Moon JY, Kim WK, Choi YH, Choi MS, Hur J, Kang JY, et al.
    Sains Malaysiana, 2017;46:217-221.
    The efficacy of the combination vaccine of the individual C-terminal fragments of ApxIA, ApxIIA and ApxIIIA of Actinobacillus pleuropneumoniae (APP) was evaluated in piglets. Twenty piglets were divided equally into 2 groups (n=10). All piglets were intramuscularly primed at 4 week-of-age (0 week post prime inoculation (WPPI)) and were intramuscularly boosted at 6 week-of-age (2 WPPI). Group A piglets were inoculated with sterile PBS and group B piglets were inoculated with the combination vaccine. Concentrations of each of the C-terminal fragment-specific IgG as determined by ELISA were significantly higher in group B than in group A from 2 WPPI until the end of this study. Clinical signs were observed from only 10% of group B piglets after the challenge with the mixture of APP serotypes 1, 2 and 5 at 4 WPPI, while 50% of group A piglets were protected against APP infections. Overall, intramuscular inoculation with the vaccine candidate can efficiently protect piglets against APP infection.
    Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
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