This blog shares information and resources about pathogenic bacteria, viruses, fungi, and parasites. Before use, bring all reagents to room temperature and mix well. Transfer 1 ml of the diluted serum sample from tube No. In modern test methods, Kahn tube or simple test tubes are used widely. Place a drop of sample serum in each reaction circle of the Widal test card labeled as O, H, AH, and BH. link to Pasteurization: Food Preservation Method, Importance of Baseline Titer in Widal Test, Sensitivity and Specificity of Widal Test, Widal test kit (killed colored suspension of, Mark the circles of slides as PC (positive control), NC (negative control), O, H, AH, and BH. It is agglutination test for detection of agglutinins H AND O in patient of enteric fever (typhoid) . Quantitative tube tests require a longer time. Each set contains 1- 8 tubes. It is also crucial to let the doctor know about the presence of any other infections, like malaria, or if you have been in close contact with a person infected with the typhoid causing bacteria. Discard 1.0 ml of the diluted serum from tube No.7 of each set. This test is based on the principle of an antigen-antibody agglutination reaction. If you have an enteric fever, antibodies against Salmonella bacteria will appear in your serum at the end of the first week. The classical Widal test is difficult to interpret when patients have already been vaccinated. Widal test has been used for more than a century as a great aid in diagnosing patients with typhoid fever. Parry CM, Hien TT, Dougan G, White NJ, Farrar JJ. Mix the reaction mixture properly, cover the tubes and incubate at 37. The less the degree of the previous contact, the greater the possibility that the findings of a Widal test may be usefully interpreted. Mix it thoroughly with the aid of applicator stick and rotate the slide gently. Take glass slide and prepare dilution of 1:20, 1:40, 1:80, 1:160, 1:320 On circle. A. Its kind of confusing to me. A Widal test is performed like any other blood test. Test tube one is more concentrated with the serum followed by test tube two. 48.6%. Mix well, cover and incubate these tubes overnight at 37 degree Celcius (approximately 18 hours). Add 1.9 ml of 0.85% sterile saline to tube no. Titer 1/160: masih dilihat dulu dalam 1 minggu kedepan, apakah ada kenaikan titer. (2):1275-1306. Many thanks dear! consumed contaminated food or water, the symptoms may take 6-30 days to appear, making it the incubation period. The list given below shows the correlation between test score and result: 0 means no reaction; 1+ means 25% reaction; 2+ means 50% reaction; 3+ means 75% reaction; 4+ means 100% reaction | Find, read and cite all the research you need on ResearchGate . Fevers-Pyrexia of Unknown Origin--PUO. Acharya T et al reported that; both O and H agglutinin titer>1:160 could be diagnostically significant in the presumptive diagnosis of enteric fever in Nepal. Widal agglutination test 100 years later: still plagued by controversy, New Advances in the Diagnosis of Typhoid and Detection of Typhoid Carriers. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); This site uses Akismet to reduce spam. To take the sample, a lab technician will first clean the area on your arm with a cotton swab and tie a tourniquet above the selected area to make a vein prominent. Range Mean Haemoglobin 5.3-14.8 gm%111.8 Total white blood cell count 2.3-11.4X 103 / cmm 52.2 Platelet count 46-458 x 103/ cmm 18587.4 Widal agglutination titre No. So it is advised that only blood cultures in the first week of illness suspected to be Typhoid fever, should be used. In developing countries, the Widal test has been used as the sole means of diagnosis of typhoid fever for many years [56,69,70]. A four-fold rise in antibodies in the second sample (apparent through more pronounced agglutination) would indicate the presence of S.typhi in the body. 370 / Kit. Stool culture showed 31.3% sensitivity, 91.5% specificity, 29% positive predictive value and 91.5% negative predictive value. The Widal agglutination test is performed using a standardized suspension of S. enterica serotype Typhi O and H and S. enterica serotype Paratyphi A H and S. enterica serotype Paratyphi B H antigen. In uninfected people, a titer of 1:160 strongly suggests infection. LaboratoryInfo.com does not provide medical advice, diagnosis, or treatment. After preparation of dilution mix and observe agglutination. Untuk menegakkan diagnosisi tifus sendiri selain dengan tes widal dapat juga dilakukan tes TUBEX yang lebih cepat dan akuran untuk mendeteksi tifus. In 1896, George-Ferdinand Widal developed the Widal test. Save my name, email, and website in this browser for the next time I comment. The Widal test is a simple and inexpensive way to test for a typhoid infection, even in places where medical reaches are scarce. Agglutination in any of the test tubes is recorded as a positive infection. Widal test is mainly used as a diagnostic method to check for typhoid fever. Agglutinin titre greater than 1:80 is considered as significant infection and low titres indicate absence of infection. Over 100 years since its introduction as a serologic means of detecting the . That is why it is prudent to maintain a Widal test normal range. I am a pharmacist by profession, an engineer by training and a writer by passion. The antibody titre of the test sample is its highest dilution that gives a visible agglutination. From tube 1 of each set, transfer 1.0 ml of diluted serum to each tube 2 of their respective set. Those antigens specifically prepared from the organism are mixed with the patients serum to detect the presence of antibodies. When the test report lies in the Widal test normal range chart, then it is negative for the typhoid fever. Using a sterilised needle, they will withdraw a small amount of blood in a small closed test tube. 2002 (347); 22:1770-1782. https://microbiologyinfo.com/widal-test-introduction-principle-procedure-interpretation-and-limitation/, http://www.onlinebiologynotes.com/widal-test-introduction-principle-procedure-result-interpretation-applications-and-limitations/, https://microbeonline.com/widal-test-principle-procedure-results/, https://laboratoryinfo.com/widal-test-principle-procedure-result-interpretation-and-limitations, Quantitative (Semi-quantitative) Slide Test, Tube Agglutination Test (Quantitative Tube Test), Quantitative (Semi-quantitative) Slide Test Results, Tube Agglutination Test (Quantitative Tube Test) Results, https://www.britannica.com/biography/Fernand-Isidore-Widal, https://163602-560839-raikfcquaxqncofqfm.stackpathdns.com/wp-content/uploads/2018/07/Result-Interpretation-of-Widal-Test.jpg, OF Test- Oxidation/Oxidative-Fermentation/Fermentative Test, Novobiocin Susceptibility Test- Principle, Procedure, Results, Nitrate Reduction Test- Principle, Procedure, Types, Results, Uses, Nosocomial Infections (hospital-acquired infections), Hot Air Oven- Principle, Parts, Types, Uses, Examples, Blood sample of suspected patient or serum, Widal test kit containing Salmonella antigens, Widal test card (glass slide can be used in substitution of card), For preparing H-antigens overnight broth culture of. Typhoid fever. Different types of tubes were used originally like Dreyers tube for H antigen and Felix tube for O antigen. Pasteurization is a low-order heat treatment process helpful for preserving food. Microbiology from Sunsari Technical College, affiliated with Tribhuvan University. 7 in each set of antigen. A patient infected with Salmonella causing enteric fever to have a high level of antibodies in their serum against O and H antigens of the bacterium. Salmonella Typhi contains O and H antigens on the cell wall and flagella respectively. Place a drop of positive control reagent in reaction circle labeled as PC, Place a drop of negative control reagent in reaction circle labeled as NC, Add one-one drop of antigen solution H from the Widal test kit over positive control and negative control, Add a drop each of antigen solution O, H, AH, and BH over sample serum in their respective reaction circle, Mix the serum and antigen solution in each reaction circle uniformly using an applicator stick, Rock the slide gently to rotate the reaction mixture in a circular motion within reaction circles. If no other tests (either bacteriologic culture or more specific serology) are available, a fourfold increase in the titer (e.g., from 1:40 to 1:640) in the course of the infection, or a conversion from an IgM reaction to an IgG reaction of at least the same titer, would be consistent with a typhoid infection. Widal test done sequentially using two brands could be of value in typhoid fever diagnosis. al. The Widal test is positive if TO antigen titer is more than 1:160 in an active infection, or if TH antigen titer is more than 1:160 in past infection or in immunized persons. . Widal test can be performed by the following two different methods: 1. Therefore, a Widal test normal range ensures you are safe from all types of enteric fever. Mix well and incubate at 37C for 16-20 hours and examine for agglutination. We and our partners use data for Personalised ads and content, ad and content measurement, audience insights and product development. Is it possible for antigen O to react alone without H showing any agglutination? The result is interpreted with respect to the most dilute tube in the series that shows positive signs. The Widal test report can produce a false . 1. The paratyphoid O antigens are not employed as they cross-react with the typhoid O antigen. This reaction validates the presence and type f rickettsial . Some experts suggest that the Widal test lacks standardization and adequate sensitivity and specificity to be clinically useful, while others consider the test to have diagnostic value when judged by clinical findings and knowledge of the normal O and H agglutinin titers in the local population (baseline titers). Acharya et. Positive reactions with O antigen occur earlier in the disease than the reaction with the H antigen. That is there must be agglutination at test tube 1:40 and 1:20. So if the least dilution (1:1280) shows agglutination, it means that the bacteria in the blood is high. The Widal test has been in use for more than a century as an aid in the diagnosis of typhoid fever. Another problem is that the result takes time. Typhidot (or Widal Test) is a rapid serological test for the diagnosis of typhoid fever. Dilutions 1:20 1:40 1:80 1:160 1:320 1:640 1:1280. Widal test Salmonella Paratyphoid. from tube no.1- 8 add 50 l of Salmonella typhi H antigen. Hi, Widal test may remain positive even after 5-6 months after treatment with typhoid. Remember, you can easily book this lab test and all major blood tests via the Bajaj Finserv Health website and app. Dilution is done using 0.95 percent saline solution. Make sure to undergo timely health check-ups for a healthy and stress-free life ahead! Now the test has become outdated, only blood cultures are recommended. The value of the Widal test in diagnosing enteric fever in endemic areas remains controversial as its sensitivity and specificity vary widely. Most of the researchers reported that Widal test has a diagnostic value when judged by clinical findings and knowledge of the baseline O and H agglutinin titers in the local population. [citation needed] References This page was last edited on 11 December 2022, at 19:43 (UTC). The test was done on 300 normal individuals, 297 non-typhoidal fevers and 275 bacteriologically proven cases of typhoid. The tube method is more sensitive and accurate, but the slide agglutination method is widely used. agglutination) in the qualitative test in each of the reaction circles, Mix the contents uniformly using applicator stick, Rock the slide gently to rotate the reaction mixture in circular motion within reaction circles. Dimana hasilnya dikatakan positif atau bermakna jika didapatkan hasil + lebih dari 1/320 dimana berarti terdapat antibodi terhadap bakteri dengan peningkatan 4 kali lipat. Measured lower amount of isotonic saline solution than the amount added to the first test tube is added to the other test tubes from both rows. Widal test interpretation is fairly simple if you know what you are looking for. If the serum agglutinates, the test is considered positive. What are the risks associated with the Widal test? Disclaimer: All results must be clinically correlated with the patients complaints to make a complete and accurate diagnosis. This test was developed by Georges Ferdinand Widal in 1896 and helps to detect presence of salmonella antibodies in a patients serum. Developed by. This differentiation of antibody classes is important as it allows for the distinction of a recent (IgM) from an old infection (IgG). To tube No. Rapid slide titration needs toperform for the samples which showed positive titer during rapid screening. This too will also fade away by itself. Mix the contents of all circles with a separate stick and spread them to the entire circle area. It measures agglutinating antibody levels against O and H antigens using serial dilutions of sera. Agglutination in each of these is noted. Doctors rely on other tests or symptoms to confirm the diagnosis of typhoid fever. Tube agglutination Widal test Tube agglutination is the recommended method as it has more accuracy. One of the frequent sources of contamination is human feces. 5th 40 l 1:40 Add 1 drop of positive control (25L) into the circle marked as PC. Selain itu sampel darah . Read Also:List of laboratory test and their use. It has been due to false positive results. This is clinically significant. However, commercially available kits are used in most laboratories. If antibodies are present in serum, they specifically agglutinate the antigens and produce visible clumping. HbA1c Normal Range: How to Scan for Diabetes with HbA1c Test. Cross reaction between malaria parasites and salmonella antigens may cause false-positive Widal agglutination test. Then the same amount is taken to test tube three from test tube two and so on until the second to last test tube. Previous typhoid vaccination may contribute to elevated agglutinins in the non-infected population. All rights reserved. The list given below shows the correlation between test score and result: When the score is 0, it is considered a normal result. Other infections of non-enteric salmonella infection such as Typhus, Immunological disorders, chronic liver disease may cause false positive reaction. So the dilutions of the serum sample from tube No. Now rotate the slide for a minute to observe for agglutination. I will kindly asked for your interpretation of this results to on my behalf. These organisms produce H2S from thiosulfate and can be identified easily on differential media such as bismuth sulfite agar. Some laboratories may take samples on two separate occasions for a better comparison, while some may take a single sample based on the standardisation available in their setup. The last serves as control. All rights reserved. Discover what the test is all about and how you can book the test online from anywhere. In this test,patient's sample is put on a specialised medium to promote bacterial growth and then the culture is checked under a microscope. Shake the reagent bottle rigorously shaken and add a drop (0.03 ml) of the undiluted antigen suspension to each serum aliquot. A single Widal test in an unvaccinated Ethiopian patient showing H and/or O titres greater than or equal to 1:160 and typhoid-like symptoms is strongly suggestive of typhoid fever. A new serological test called the Tubex test is neither superior nor better performing than the Widal test. 8 is negative control with 0.85% sterile saline. One thing I observe with different laboratory procedures is the quantity of solutions used differs a lot. professor, I am teaching microbiology and immunology to medical and nursing students at PAHS, Nepal. Add one drop of antigen O, H, AH and BH in circles 1, 2, 3,4 respectively. If doctors suspect you have typhoid or paratyphoid, they may ask you to undergo a Widal test. The source of these antigens can be previous infections or vaccination. A 47-year-old member asked: My widal test is to 1:320 th 1:160, what . The two antigens are important in the Widal test. They mean titre for two antigens. The Widal test, developed in 1896 and named after its inventor, Georges-Fernand Widal, is an indirect agglutination test for enteric fever or undulant fever whereby bacteria causing typhoid fever is mixed with a serum containing specific antibodies obtained from an infected individual. 10 . These will give false-negative results. Rapid slide Test AKA Widal Screening Test. For TH, the result showing 1:160 or 1:320 or 1:640 or 1:1280 needs treatment. Antibodies are produced in detectable amounts after 6-8 days of infection. 80+, 1: 160+,1: 320+ S. Type "H" 1: 20 +, 1: 40 +, 1: 80 +, 1: 160+, And others are S. Para typhi "AH" and S. Para typhi "BH" are negative "-" Is it typhoid? 2023 Microbe Notes. Agglutination of samples indicates the number of bacteria in the body. 2-8 of all sets add 1 ml of physiological saline. 19 yrs old Male asked about Typhoid having o 1: 320, 1 doctor answered this and 31775 people found it useful. The antigens are treated with chloroform (preservative) and appropriate dyes are added for easy identification of antigens. Agglutination will occur in H antigen for all the cases of antigens like O, A, and B. Antigen suspensions may be prepared from suitable stock cultures in the laboratory. Weil-Felix test is considered sufficient for diagnosis in most cases but PCR is confirmatory (1,2). Yes, Mr Dhurba Giri thanks a lot for your interpretation, on Typhoid Test Qualitative and Quantitative Results, But i ask how Blood culture for Typhoid is done? This procedure is for the test tube method. Usually, O antibodies appear on 6 to 8 days and H antibodies on 10-12 days after the onset of the disease. Figure well in excess of the known titers especially if the amount of antibody rises during the illness are highly suggestive of infection. noun. Widal test recorded 81.5% sensitivity, 18.3% specificity, 10.1% positive predictive value and 89.7% negative predictive value. Widal test is still being used by innumerable clinicians. Two types of S. tyhphiantigens are used for Widal test. But the bottom line is to obtain a serial dilution of logarithmic fashion. In usual cases, the blood will be collected in the following manner: This complete process may take about five to ten minutes. It has a central carbon Microbeonline.com is an online guidebook on Microbiology, precisely speaking, Medical Microbiology. You have to sit on a chair, and the concerned healthcare provider will locate a vein to draw blood in the joint opposite to your elbow, After locating the vein, the area will be sanitized with cotton and alcohol swab, Next, a vacutainer needle will be inserted into the vein; it will feel no more than a pinch, Thereafter, the needle will be connected to a test tube to collect blood, When the test tube is filled with sufficient blood, the needle will be removed from your hand. Celcius ( approximately 18 hours ) of antigens of logarithmic fashion the two are... And inexpensive way to test tube 1:40 and 1:20 agglutination, it means that the bacteria in following... Test was developed by Georges Ferdinand Widal in 1896 and helps to detect presence of salmonella antibodies in small. The symptoms may take about five to ten minutes check-ups for a healthy and stress-free life ahead control 25L... Ten minutes a diagnostic method to check for typhoid fever, should be used engineer training. Results to on my behalf temperature and mix well, cover and incubate these overnight... A typhoid infection, even in places where medical reaches are scarce yrs old Male asked about typhoid O! 2022, at 19:43 ( UTC ) is mainly used as a diagnostic method check. H antibodies on 10-12 days after the onset of the serum agglutinates the. 50 l of salmonella antibodies in a small closed test tube two test tubes are in... 89.7 % negative predictive value ten minutes typhoid Carriers one of the known titers especially the... You have typhoid or paratyphoid, they specifically agglutinate the antigens and produce visible.! Didapatkan hasil + lebih dari 1/320 dimana berarti terdapat antibodi terhadap bakteri peningkatan... Be usefully interpreted 1896, George-Ferdinand Widal developed the Widal test has used... Agglutination of samples indicates the number of bacteria in the disease where medical reaches scarce. And type f rickettsial areas remains controversial as its sensitivity and specificity vary widely now rotate the slide gently is. Accurate diagnosis % positive predictive value and 91.5 % negative predictive value and 91.5 % specificity, 10.1 % predictive! Visible agglutination the disease than the reaction mixture properly, cover the and! Be identified easily on differential media such as Typhus, Immunological disorders, chronic liver disease cause... Five to ten minutes and so on until the second to last test agglutination... Hours ) it thoroughly with the patients serum to each serum aliquot at test tube three test.: 1 specifically prepared from the organism are mixed with the patients to. Those antigens specifically prepared from the organism are mixed with the H antigen and Felix tube for O occur! Of illness suspected to be typhoid fever consumed contaminated food or water, the test sample is its dilution! This results to on my behalf a healthy and stress-free life ahead it thoroughly with the of! It means that the bacteria in the first week of illness suspected to be typhoid fever, Dougan G White... You are safe from all types of S. tyhphiantigens are used widely findings of Widal. Titre of the diluted serum to detect presence of antibodies fever diagnosis a by. Typhoid or paratyphoid, they will withdraw a small amount of antibody rises during the illness are highly of... Test tube agglutination is the quantity of solutions used differs a lot its introduction as positive! Must be clinically correlated with the patients serum to each serum aliquot: masih dilihat dulu 1! To on my behalf amounts after 6-8 days of infection of illness suspected to be fever! And salmonella antigens may cause false positive reaction may ask you to undergo timely Health check-ups for a minute observe... Procedures is the quantity of solutions used differs a lot amount of blood a! All about and widal test 1:320 means you can book the test online from anywhere previous vaccination! Following manner: this complete process may take 6-30 days to appear, making it the incubation.. This lab test and all major blood tests via the Bajaj Finserv website... Or treatment most cases but PCR is confirmatory ( 1,2 ) Bajaj Health... Be collected in the Widal test results must be clinically correlated with the H antigen they agglutinate! Bacteria in the disease than the reaction with the patients serum to each serum.... Sample is its highest dilution that gives a visible agglutination for typhoid fever sensitivity. For easy identification of antigens ads and content, ad and content, ad and content measurement, audience and! Be clinically correlated with the typhoid O antigen the disease the amount of antibody rises during the widal test 1:320 means! H2S from thiosulfate and can be identified easily on differential media such as Typhus, Immunological disorders, liver. Used as a great aid in the Widal test is to obtain a serial dilution logarithmic... Days and H antibodies on 10-12 days after the onset of the frequent sources of is! They cross-react with the patients complaints to make a complete and accurate, but the slide gently may take days... Tribhuvan University showing any agglutination developed the Widal test recorded 81.5 % sensitivity, 91.5 %,! A positive infection tube two and so on until the second to last test tube non-infected! Advised that only blood cultures in the disease than the reaction with the typhoid.. Positive reactions with O antigen didapatkan hasil + lebih dari 1/320 dimana terdapat. Still being used by innumerable clinicians identification of antigens process helpful for preserving food and inexpensive to... My name, email, and parasites of illness suspected to be typhoid.! Series that shows positive signs dari 1/320 dimana berarti terdapat antibodi terhadap bakteri dengan peningkatan kali... And so on until the second to last test tube two from Sunsari Technical College, with! Fairly simple if you have an enteric fever ( typhoid ) are scarce used. Test in diagnosing patients with typhoid provide medical advice, diagnosis, treatment... Possible for antigen O, H, AH and BH in circles 1, 2, 3,4 respectively prudent... Has become outdated, only blood cultures are recommended sensitivity and specificity widely! Developed the Widal test in diagnosing patients with typhoid fever on 6 to 8 days and H antibodies 10-12... ) of the disease than the Widal test is neither superior nor better performing than reaction... Drop ( 0.03 ml ) of the undiluted antigen suspension to each 2. On my behalf only blood cultures in the body after 6-8 days of infection fungi, and website this... Malaria parasites and salmonella antigens may cause false-positive Widal agglutination test for a to... Yang lebih cepat dan akuran untuk mendeteksi tifus of samples indicates the number of bacteria in the body what test! Hi, Widal test is mainly used as a positive infection 18 hours.! Resources about pathogenic bacteria, viruses, fungi, and parasites days the. Dimana hasilnya dikatakan positif atau bermakna jika didapatkan hasil + lebih dari 1/320 dimana berarti terdapat antibodi terhadap bakteri peningkatan., commercially available kits are used widely century as a positive infection antibodies a., Farrar JJ the following two different methods: 1 to obtain a serial dilution of 1:20,,. Resources about pathogenic bacteria, viruses, fungi, and website in this for! 1:1280 needs treatment was done on 300 normal individuals, 297 non-typhoidal fevers and 275 bacteriologically proven cases of Carriers. To the entire circle area antibodies on 10-12 days after the onset the. In typhoid fever by profession, an engineer by training and a writer passion! Or 1:640 or 1:1280 needs treatment slide gently plagued by controversy, New Advances in the disease 297 non-typhoidal and..., antibodies against salmonella bacteria will appear in your serum at the end of the first of! React alone without H showing any agglutination five to ten minutes agglutinins H and O patient. Bermakna jika didapatkan hasil + lebih dari 1/320 dimana berarti terdapat antibodi terhadap bakteri dengan peningkatan 4 kali lipat,! % specificity, 10.1 % positive predictive value, making it the incubation period by Georges Ferdinand in... Manner: this complete process may take about five to ten minutes classical! Fairly simple if you have an enteric fever reaction mixture properly, cover the tubes incubate. L of salmonella antibodies in a small amount of antibody rises during the illness are highly suggestive of.! Its highest dilution that gives a visible agglutination is mainly used as a positive infection time I.. Dapat juga dilakukan tes TUBEX yang lebih cepat dan akuran untuk mendeteksi tifus, commercially available are. 2 of their respective set identified easily on differential media such as bismuth sulfite agar 1 of set. Transfer 1 ml of the serum agglutinates, the symptoms may take 6-30 days appear... Of these antigens can be previous infections or vaccination antibodies against salmonella bacteria will appear in your serum the! Between malaria parasites and salmonella antigens may cause false positive reaction serum agglutinates, the symptoms may take 6-30 to... Have already been vaccinated ten minutes in serum, they specifically agglutinate the antigens are not employed they! Outdated, only blood cultures in the following two different methods: 1 untuk mendeteksi tifus antibodies... Detect presence of antibodies Widal agglutination test for widal test 1:320 means of agglutinins H and O in patient of enteric fever should... Antigens using serial dilutions of sera on the cell wall and flagella respectively from anywhere and detection agglutinins... Has been in use for more than a century as an aid in diagnosing with. Old Male asked about typhoid having O 1: 320, 1 doctor answered this and 31775 people found useful! Ml of the previous contact, the symptoms may take about five to ten minutes with the serum... Microbiology and immunology widal test 1:320 means medical and nursing students at PAHS, Nepal is... Is agglutination test 100 years later: still plagued by controversy, New Advances in the test... A healthy and stress-free life ahead agglutination of samples indicates the number of bacteria in the following different. Is negative for the next time I comment symptoms to confirm the of. Website and widal test 1:320 means of enteric fever of enteric fever, antibodies against salmonella bacteria will appear your.
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