Mantar enfeksiyonlarının serolojik tanısı

İnvaziv mantar hastalıklarının tanısında, günümüzde kullanılan yöntemlerin duyarlılık ve özgüllüğündeki yetersizlikler veya klinik olarak yararlı olabilecek bir sonucun elde edilmesinin çok zaman alması nedeniyle zorluklar yaşanmaktadır. Hasta sonuçlarında, hızlı tanı anahtar noktadır. Kültür testleri dışında, konaktaki özgül antikor yanıtının ölçülmesine dayalı serolojik testlerin, hızlı uygulanabilmesi ve invaziv örnek alım yöntemleri gerektirmemesi bu testleri çekici kılmaktadır. Ancak özellikle immünsupresif durumlar nedeniyle, özgül antikor yanıtı oluşturamayan hastalarda bu testler her zaman invaziv hastalık belirteci olmamaktadır. Mikrobiyal antijenlerin tespitinin ise genellikle daha fazla miktarda mikrobiyal yük gerektirmesi duyarlılığını kısıtlamaktadır. Yine de çok sayıda başarıyla çalışan antijen tespit sistemleri geliştirilmiştir ve bunların bazıları halen yaygın olarak kullanılmaktadır. Dolaşımdaki Aspergillus galaktomannanlarını tespit için sıçan monoklonal EBA2'leri kullanan emzim immünoassay yöntemi (Platelia) ticari olarak kullanımdadır. Bu test 0,5-1,0 ng/ml düzeyindeki galaktomannanı tespit edebilmektedir. Ayrıca, galaktomannan klinik belirti ve semptomlar oluşmadan invaziv aspergillozun erken safhalarında serumda tespit edilebilmektedir. Ancak, bu testin yanlış pozitif veya negatif sonuçlar gibi dezavantajları bulunmaktadır. (1,3)-?-D-glukan Zygomycetes'ler hariç mantar hücre duvarının karakteristik bir birleşenidir. İnvaziv enfeksiyonlar sırasında Aspergillus, Candida, Fusarium, Trichosporon, Saccharomyces, Acremonium ve Pneumocyctis jirovecii içeren birçok mantar tarafından (1,3)-?-D-glukan kana salınmaktadır. (1,3)-?-D-glukan konsantrasyonunun belirlenmesinde kullanılan yöntem, atnalı (horsecrab shoe) yengecinin koagülasyon faktörü olan faktör G'nin glukanı aktifleştirmesine dayanmaktadır. Hem Avrupa'da hem de ABD'de (1,3)-?-D-glukan için çeşitli ticari kitler kullanımdadır. Mannan invaziv kandidiyazisi bulunan hastalarda dolaşımdaki ana antijendir. Mannana karşı oluşan antikorların tespiti için birçok serolojik test bulunmaktadır. Ancak bu testler, kolonizasyon ve invaziv enfeksiyon ayrımında başarısız olmaktadır. Mannan antijeniyle kombine edilip çalışıldığında, duyarlılık %80 ve özgüllük %93 olmaktadır. Kriptokokkal enfeksiyonların tespitinde serebrospinal sıvıdan veya serumdan kapsüler antijen tespiti kullanılmaktadır. Kapsüler antijen tespiti için hem lateks aglütinasyon hem de enzimimmünoassay testleri bulunmaktadır. Mantar enfeksiyonlarının tanısı için son yıllarda hızla ilerleme kaydedilerek, mantar enfeksiyonlarının erken tespitini sağlayan birçok yeni yöntem geliştirilmiştir. Ancak en iyi yaklaşımın kültür veya mikroskopi gibi konvansiyonel yöntemlerle serolojik yöntemlerin kombine edilmesi olduğu düşünülmektedir.

Serological diagnosis of fungal infections

Difficulties are being faced in the diagnosis of invasive fungal diseases due to the lack of sensitivity and specificity of the current diagnostic methods or taking too long time to get a result having clinical importance. Rapid diagnosis is the key point in patient outcomes. Excluding the culture tests, because of its rapid application and no need for invasive sampling, was making the serological tests based on the measurement of response of specific host antibody became attractive. However especially due to immünosuppressive conditions, these tests are not always the indicator of invasive disease in the patients who have no ability to produce specific antibody response. Detection of microbial antigens generally requiring a relatively large microbial burden, limits the assay sensitivity. Nonetheless, several examples of successful antigen detection systems have been developed, and some of these are widely used. To detect circulating Aspergillus galactomannans enyzme immünoassay (Platelia) method utilizing the rat monoclonal antibody EB-A2 is commercially in use. This test is able to detect galactomannan in the level of 0.5-1.0 ng/ml. Furthermore, galactomannan could be detected in the serum at a early stage of invasive aspergillosis before clinical signs and symptoms occured. However, this test has some disavantages like false positive or negative results. (1.3)-β-D-glucan is a characteristic cell-wall component of fungi except for Zygomycetes. A broad range of fungi including Aspergillus, During invasive infections (1.3)-β-D-glucan is released to the blood by some fungi containing Aspergillus, Candida, Fusarium, Trichosporon, Saccharomyces, Acremonium and Pneumocyctis jirovecii. The method used for determination of (1.3)-β-D-glucan concentration depends on activation of glucan by factor G, a horsecrab shoe coagulation factor. There are some commercial test kits available for (1.3)-β-D-glucan both in Europe and USA.Mannan is the major circulating antigen in patients who have invasive candidiasis. There are several serological tests for the detection of antibodies formed against mannan. However, these tests are usually failed in discriminating between colonization and invasive infection. When combination with mannan antigen test was performed, it gave sensivity of 80% and specifity of 93%. In the diagnosis of cryptococcal infections detection of capsular antigen in the both cerebro-spinal fluid or serum, is used. Both latex aglutination and enzymeimmünoassay tests are found to detection of capsular antigen. The diagnosis of fungal infections has moved forward in the past few years and there are many newly developed methods that are likely to provide early detection of fungal infections. However, it seems that best approach is combining both traditional methods like culture or microscopy with serological methods

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