Investigation of the presence of encephalitozoon intestinalis and enterocytozoon bieneusi in bone marrow transplant patients by IFA-MAbs method

Mikrosporidian patojenler her yerde bulunabilen, omurgalı ve omurgasız konaklarda enfeksiyona neden olabilen, 144 cins altında 1200'den fazla türü tanımlanmış zorunlu hücre içi ökaryotik parazitlerdir. İnsanlarda patojen olarak tanımlanan 14 microsporidia türü arasında en sık saptananlar Enterocytozoon bieneusi ve Encephalitozoon intestinalis olup gastrointestinal sistemde enfeksiyona yol açmaktadırlar. Bu etkenler, özellikle immün sistemi baskılanmış hastalarda kronik ishallerin yanı sıra, yaygın enfeksiyonlara neden olarak hayatı tehdit eden ağır klinik tablolar oluşturmaktadır. Microsporidia sporları çok küçük olduklarından, rutin dışkı incelemelerinde genellikle gözden kaçmaktadır. Bu nedenle tanıda moleküler yöntemler ve mümkünse transmisyon elektron mikroskopi altın standart olarak kullanılmaktadır. Bu yöntemlerin uygulanamadığı laboratuvarlarda ise, monoklonal antikorların kullanıldığı immünofl oresan antikor (IFA-MAbs) yöntemi, konvansiyonel yöntemlere üstünlüğü nedeniyle tercih edilebilir. Bu çalışmada, kemik iliği transplant (KİT) hastalarında E.intestinalis ve E.bieneusi varlığının IFA-MAbs yöntemi ile araştırılması amaçlanmıştır. Çalışmaya, 147'sinde ishal şikayeti olan 200 KİT hastası (134'ü erkek, 66'sı kadın; yaş ortalaması: 43.2 ± 15.01 yıl) ile kontrol olarak 80 sağlıklı birey (43'ü erkek, 37'si kadın; yaş ortalaması: 31.9 ± 11.76 yıl) dahil edilmiştir. Tüm olguların dışkı örnekleri, konvansiyonel yöntemlerin (nativ-lugol ve modifi ye aside dirençli boyama) yanı sıra, ticari bir IFA-MAbs (Bordier Affi nity Products, İsviçre) yöntemiyle de değerlendirilmiştir. Hastaların %25.5 (51/200)'inde E.intestinalis, %4 (8/200)'ünde E.bieneusi ve %9.5 (19/200)'inde her ikisi birden olmak üzere, toplam %39 (78/200)'unda pozitifl ik saptanmıştır. Bu oranlar kontrol grubu için sırasıyla, %5 (4/80), %2.5 (2/80), %3.8 (3/80) ve %11.3 (9/80) olarak tespit edilmiştir. Hasta ve kontrol gruplarında saptanan pozitifl ik oranları arasındaki fark istatistiksel olarak anlamlı bulunmuştur (%39'a karşı %11.3, p< 0.05). Hasta grubunda pozitif bulunan 78 hastanın 67 (%85.9)'sinin ishalli olduğu izlenmiş; ishal varlığı ile parazit pozitifl iği arasında istatistiksel olarak anlamlı bir ilişkinin olduğu görülmüştür (p< 0.05). Sonuç olarak, KİT hastalarının yaşam kalitesini yükseltmek ve tedavi sürecindeki olumsuzlukları azaltmak için, bu hastaların, özellikle de gastrointestinal yakınması olanların düzenli aralıklarla E.intestinalis ve E.bieneusi açısından değerlendirilmesi gerektiği düşünülmüştürMicrosporidian pathogens are obligatory intracellular eukaryotic parasites which can be found worldwide. They have been represented in 144 genera and more than 1200 species that may cause infections in both vertebrate and invertebrate hosts. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species among 14 species of microsporidia identifi ed as human pathogens and they cause infections in the gastrointestinal tract. These species may also cause chronic diarrhea particularly in immunocompromised patients, as well as disseminated infections with severe clinical conditions which can be life-threatening. Since the spores of microsporidia are quite small-sized structures, they frequently may be overlooked in routine stool examinations. Therefore, molecular methods and transmission electron microscopy, if possible, are used as the gold standard methods in laboratory diagnosis. In laboratories in which those methods could not be applied, immunofl uorescence assay using monoclonal antibodies (IFA-MAbs) may be advantageous compared to conventional methods. The aim of this study was to investigate the presence of E.intestinalis and E.bieneusi in bone marrow transplant (BMT) patients by using IFA-MAbs method. A total of 200 BMT patients (134 male, 66 female; mean age: 43.2 &plusmn; 15.01 years), of them 147 with diarrhea and 80 healthy subjects (43 male, 37 female; mean age: 31.9 &plusmn; 11.76 years) as control group were included in the study. All of the stool samples were examined by a commercial IFA-MAbs (Bordier Affi nity Products, Switzerland) method as well as conventional (native-lugol and modifi ed acid-fast staining) methods. Of the patients 25.5% (51/200) were positive for E.intestinalis, 4% (8/200) for E.bieneusi and 9.5% (19/200) for both of them, giving a total positivity rate of 39% (78/200). Those rates were 5% (4/80), 2.5% (2/80), 3.8% (3/80) and 11.3% (9/80), respectively for control group. The difference between the patient and control groups in terms of positivity was found statistically signifi cant (39% vs 11.3%, p&lt; 0.05). Among 78 positive BMT patients, 67 (85.9%) were suffering from diarrhea. The correlation between the presence of diarrhea and the presence of microsporidia was statistically signifi cant (p&lt; 0.05). It was concluded that, BMT patients particularly those with gastrointestinal complaints, have to be evaluated for microsporidian pathogens regularly to improve quality of life and to decrease the problems during the treatment perio

Encephalitozoon intestinalis: A Rare Cause of Diarrhea in an Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) Recipient Complicated by Albendazole-Related Hepatotoxicity

Abstract: A 50-year-old male patient previously diagnosed with acute myelomonocytic (M4) leukemia in July 2009 underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). During the pre-transplant period complete blood count (CBC), liver and renal function tests, coagulation tests, and other parameters were normal. On the first day of transplantation teicoplanin (400 mg d -1 for the first 3 d, and then 400 mg d -1 ) and caspofungin (first dose was 1×70 mg d -1 , followed by 1×50 mg d -1 ) were started intravenously due to white plaques and oropharyngeal candidiasis in the patient&apos;s mouth and perianal erythema. On the 14 th d of transplantation watery diarrhea occurred, along with abdominal discomfort, nausea, and fatigue

Investigation of the Presence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in Bone Marrow Transplant Patients by IFA-MAbs Method

Microsporidian pathogens are obligatory intracellular eukaryotic parasites which can be found worldwide. They have been represented in 144 genera and more than 1200 species that may cause infections in both vertebrate and invertebrate hosts. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species among 14 species of microsporidia identified as human pathogens and they cause infections in the gastrointestinal tract. These species may also cause chronic diarrhea particularly in immunocompromised patients, as well as disseminated infections with severe clinical conditions which can be life-threatening. Since the spores of microsporidia are quite small-sized structures, they frequently may be overlooked in routine stool examinations. Therefore, molecular methods and transmission electron microscopy, if possible, are used as the gold standard methods in laboratory diagnosis. In laboratories in which those methods could not be applied, immunofluorescence assay using monoclonal antibodies (IFA-MAbs) may be advantageous compared to conventional methods. The aim of this study was to investigate the presence of Eintestinalis and Ebieneusi in bone marrow transplant (BMT) patients by using IFA-MAbs method. A total of 200 BMT patients (134 male, 66 female; mean age: 43.2 15.01 years), of them 147 with diarrhea and 80 healthy subjects (43 male, 37 female; mean age: 31.9 11.76 years) as control group were included in the study. All of the stool samples were examined by a commercial IFA-MAbs (Bordier Affinity Products, Switzerland) method as well as conventional (native-lugol and modified acid-fast staining) methods. Of the patients 25.5% (51/200) were positive for Eintestinalis, 4% (8/200) for E.bieneusi and 9.5% (19/200) for both of them, giving a total positivity rate of 39% (78/200). Those rates were 5% (4/80), 2.5% (2/80), 3.8% (3/80) and 11.3% (9/80), respectively for control group. The difference between the patient and control groups in terms of positivity was found statistically significant (39% vs 11.3%, p< 0.05). Among 78 positive BMT patients, 67(85.9%) were suffering from diarrhea. The correlation between the presence of diarrhea and the presence of microsporidia was statistically significant (p< 0.05). It was concluded that, BMT patients particularly those with gastrointestinal complaints, have to be evaluated for microsporidian pathogens regularly to improve quality of life and to decrease the problems during the treatment period

Investigation of Epstein Barr virus serology and DNA in bone marrow transplant recipients

The aim of this study was to investigate Epstein-Barr virus (EBV) serology and EBV DNA in the patients with bone marrow transplantation. In the current study, 128 patients with bone marrow transplantation (BMT) were included in Hematology-Oncology Department of Medical Faculty, Erciyes University between June 2005 and October 2009. In order to evaluate EBV serology and DNA, 357 samples obtained from the patients were studied with enzyme-linked immunosorbent assay and real-time polymerase chain reaction (PCR). The components of EBV serology were anti-Ebstein-Barr nuclear antigen (EBNA)-1 IgG, anti-viral capsid antigen (VCA) IgG and IgM. EBV seropositivity was found in 98.4% of the samples. EBV DNA positivity was detected in 13.3% of the patients. High viral load was only seen in one patient among all the EBV DNA positivity. No patient showed any symptoms regarding EBV virus and admitted with post-transplant lymphoproliferative disease. In conclusion, for the diagnosis and follow up of EBV infections in patients with BMT, detection of EBV DNA by PCR method is also beneficial in addition to serological tests regarding EBV

Prevalence of encephalitozoon intestinalis and enterocytozoon bieneusi in cancer patients under chemotherapy

Zorunlu hücre içi parazitler olan mikrosporidia türleri, özellikle immün sistemi baskılanmış hastalarda ciddi enfeksiyonlara yol açan en önemli fırsatçı patojenlerdendir. İnsanı enfekte eden 14 mikrosporidia türü arasında Enterocytozoon bieneusi ve Encephalitozoon intestinalis en yaygın olanlarıdır. Bu çalışmada, kemoterapi alan kanserli hastalarda E.intestinalis ve E.bieneusi prevalansının, immünofloresan antikor ve konvansiyonel boyama yöntemleri ile araştırılması amaçlanmıştır. Çalışmaya, hastanemizin onkoloji ve hematoloji kliniklerinde takip edilen 93 kanserli hasta (58 erkek, 35 kadın) ile kontrol olarak 30 (13 erkek, 17 kadın) sağlıklı gönüllüden alınan toplam 123 dışkı örneği dahil edilmiştir. Hastaların 51 (%55)’inde ishal şikayeti mevcuttur. Tüm örneklerde E.intestinalis ve E.bieneusi varlığı, monoklonal antikorların kullanıldığı ticari bir immünofloresan antikor yöntemi (IFA-MAb; Bordier Affinity Products, İsviçre) ile araştırılmış; ayrıca 50 örnek modifiye trikrom, aside dirençli trikrom ve kalkoflor boyama yöntemleri ile de incelenmiştir. IFA-MAb yöntemi ile hastaların 43 (%46.2)’ünde E.intestinalis, 9 (%9.7)’unda E.bieneusi ve 13 (%14)’ünde karışık enfeksiyon olmak üzere toplam 65 (%69.9) olguda pozitiflik saptanmış; kon- trol grubunda ise 2 (%6.7) E.intestinalis, 1 (%3.3) E.bieneusi ve 2 (%6.7) karışık enfeksiyon olmak üzere toplam 5 (%16.7) pozitif sonuç alınmıştır. Hasta ve kontrol grubunun pozitiflik oranları arasındaki fark istatistiksel olarak anlamlı bulunmuştur (p< 0.05). İshalli hastalardan %68.6 (35/51)’sının mikrosporidia ile enfekte olduğu izlenmiş; microsporidia pozitiflik oranı ishali olan ve olmayan (%48.6) olgular arasında anlamlı fark göstermiştir (p< 0.05). Tüm yöntemlerin birlikte uygulandığı 50 örnek değerlendirildiğinde; microsporidia pozitiflik oranları IFA-MAb yöntemi ile %66 (n= 33), modifiye trikrom boyama ile %34 (n= 17), aside dirençli trikrom boyama ile %24 (n= 12) ve kalkoflor boyama ile %42 (n= 21) olarak belirlenmiştir. Bu veriler, mikrosporidia tanısında konvansiyonel boyama yöntemleri ile birlikte IFA-MAb yönteminin kullanılmasının duyarlılığı artıracağını göstermektedir. Sonuç olarak çalışmamızda, kemoterapi alan kanserli hastalarda E.intestinalis ve E.bieneusi prevalansının oldukça yüksek (%69.9) olduğu saptanmış; bu hastaların düzenli olarak mikrosporidian patojenler açısından taranmasının uygun olacağı düşünülmüştür.Microsporidia species are obligate intracellular parasites and constitute one of the most important opportunistic pathogens that can cause severe infections especially in immunocompromised patients. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species among 14 microsporidia species identified as human pathogens. The aim of this study was to investigate the prevalence of E.intestinalis and E.bieneusi in cancer patients under chemotherapy by immunofluorescent antibody and conventional staining methods. A total of 123 stool samples obtained from 93 patients (58 male, 35 female) with cancer who were followed in oncology and hematology clinics of our hospital and 30 healthy volunteers (13 male, 17 female) were included in the study. Fifty-one (55%) of the patients had complain of diarrhea. The presence of E.intestinalis and E.bieneusi were investigated by a commercial immunofluorescence antibody test using monoclonal antibodies (IFA-MAbs; Bordier Affinity Products, Switzerland) in all of the samples, and 50 of the samples were also investigated by modified trichrome, acid-fast trichrome and calcofluor staining methods. A total of 65 (69.9%) patients were found positive with IFA-MAbs method, including 43 (46.2%) E.intestinalis, 9 (9.7%) E.bieneusi and 13 (14%) mixed infections. In the control group, 5 (16.7%) subjects were positive with IFA-MAbs method, including 2 (6.7%) E.intestinalis, 1 (3.3%) E.bieneusi and 2 (6.7%) mixed infections. The difference between the positivity rate of the patient and control groups was statistically significant (p&lt; 0.05). Of the patients with diarrhea, 68.6% (35/51) were infected with microsporidia, and the difference between cases with and without (48.6%) diarrhea was statistically significant (p&lt; 0.05). When 50 samples in which all of the methods could be performed were evaluated, the frequency of microsporidia were detected as follows; 66% (n= 33) with IFA-MAbs, 34% (n= 17) with modified trichrome staining, 24% (n= 12) with acid-fast trichrome staining and 42% (n= 21) with calcofluor staining methods. Our data indicated that the use of IFA-MAbs method along with the conventional staining methods in diagnosis of microsporidia will increase the sensitivity. As a conclusion, the prevalence of E.intestinalis and E.bieneusi in cancer patients under chemotherapy was detected quite high (69.9%) in our study, it would be appropriate to screen these patients regularly in terms of microsporidian pathogens