Candida albicans suşlarının proteinaz enzim aktivitelerinin araştırılması ve çeşitli olgu gruplarında proteinaz enzimine karşı serumda antikor aranması
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Date
1999
Authors
Akçağlar, Sevim
Journal Title
Journal ISSN
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Publisher
Uludağ Üniversitesi
Abstract
Bu çalışmada 15 tanesi kolonizasyon örneği olarak boğaz sürüntülerinden, 15 tanesi etken olarak kan örneklerinden izole edilen toplam 30 Candida albicans suşlarının proteinaz aktivitelerine ve saflaştırılan proteinaz enziminin çeşitli çalışma gruplardaki serodiyagnostik tanı değerine bakıldı. Proteinaz aktiviteleri spektrofotometrik ve plak yöntemi olmak üzere farklı iki yöntemle ölçüldü. Proteinaz enziminin serodiyagnostik tanıdaki yerini belirlemek için antijen olarak kullanılmak üzere saflaştırılması, anyon değişim kromatografisi yöntemi ile standart C albicans ATCC (10231) susundan yapıldı. Kolonizasyon grubu ve kandidemi grubuna ek olarak 50 erkek kan donöründen oluşan kontrol grubuyla beraber toplam 80 serumda proteinaz antikorları ELISA yöntemiyle arandı. Proteinaz aktivitesi yönünden her iki yöntemle de etken ve kolonizasyon gruplarından elde edilen suşların farklı olmadığı görüldü (p> 0.05). Antikor taramalarında ise, en düşük antikor seviyeleri beklenildiği gibi kontrol grubunda (ort: 0.116 ± 0.020) tesbit edilse de üç grup arasındaki fark anlamlı bulunmadı (p>0.05). Ancak gruplar arasında ikili yapılan karşılaştırmalarda kontrol grubuyla kolonizasyon grubu arasındaki antikor seviyeleri anlamlı olarak farklıydı (p<0.01). Bu bulgulara göre in vitro stabil ortamlarda proteinaz enzim aktivitesini ölçmenin C albicans suşlarında ayırıcı olamayacağı; proteinaz enzimine karşı antikor taramalarının da, olasılıkla enzimin adezyondan da sorumlu olması nedeniyle invazif enfeksiyonları tanıyamayacağı; dolayısıyla invazif kandida enfeksiyonlannda kanda ve dokularda antijen tarama yöntemlerinin diyagnostik açıdan daha değerli olduğu kanısına varıldı.du.tr/submit
In this study proteinase enzyme activities of 30 Candida albicans, which were isolated from 15 colonization samples from throat swabs and 15 infection agent from blood samples, were measured and serodiagnostic value of purified enzyme was evaluated. Spectrophotometric and plate methods were used to determine proteinase activity. Anion exchange chromatography was used to purify standart C albicans ATCC (10231) strain for the estimation of usefulness of enzyme antigen as a serodiagnostic parameter. In addition, a total of 80 serum samples collected from 50 male blood donors without colonization (control grup), from patient group with colonization and from candidemia were examined by ELISA for the antibody againts purified proteinase. It has been shown that in terms of proteinase activitiy there was no statistically significant differences between strains obtained from infection and colonization sites by both methods (p>0.05). As expected the lowest antibody levels were found from the serum of control grup (mean 0.116 ± 0.020), but there was no significant differences between different study grups (p>0.05). Instead, when comparison have been done between two groups, from the serum of colonization significant antibody levels were obtained than control serums (p<0.01 ). According to these findings, at in vitro conditions proteinase enzyme activities cannot delineate C albicans strains. Antibodies against purified proteinase enzyme cannot be used to diagnose invasive Candida infections because of the role of enzyme in adhesion process to mucosal surfaces as well. So, it is believed that proteinase enzyme can be more diagnostic for invasive infections when searchs as an antigen from the blood and tissue samples.
In this study proteinase enzyme activities of 30 Candida albicans, which were isolated from 15 colonization samples from throat swabs and 15 infection agent from blood samples, were measured and serodiagnostic value of purified enzyme was evaluated. Spectrophotometric and plate methods were used to determine proteinase activity. Anion exchange chromatography was used to purify standart C albicans ATCC (10231) strain for the estimation of usefulness of enzyme antigen as a serodiagnostic parameter. In addition, a total of 80 serum samples collected from 50 male blood donors without colonization (control grup), from patient group with colonization and from candidemia were examined by ELISA for the antibody againts purified proteinase. It has been shown that in terms of proteinase activitiy there was no statistically significant differences between strains obtained from infection and colonization sites by both methods (p>0.05). As expected the lowest antibody levels were found from the serum of control grup (mean 0.116 ± 0.020), but there was no significant differences between different study grups (p>0.05). Instead, when comparison have been done between two groups, from the serum of colonization significant antibody levels were obtained than control serums (p<0.01 ). According to these findings, at in vitro conditions proteinase enzyme activities cannot delineate C albicans strains. Antibodies against purified proteinase enzyme cannot be used to diagnose invasive Candida infections because of the role of enzyme in adhesion process to mucosal surfaces as well. So, it is believed that proteinase enzyme can be more diagnostic for invasive infections when searchs as an antigen from the blood and tissue samples.
Description
Keywords
Candida albicans, Enzymes, Enzimler
Citation
Açağlar, S. (1999). Candida albicans suşlarının proteinaz enzim aktivitelerinin araştırılması ve çeşitli olgu gruplarında proteinaz enzimine karşı serumda antikor aranması. Yayınlanmamış doktora tezi. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.