Publication: Amebiyaz’ın laboratuvar tanısında kültür, direkt incelenme ve antijen arama yöntemlerinin karşılaştırılması
Abstract
Entamoeba histolytica infeksiyonlarının en sıklıkla karşılaşılan klinik formu, barsak amebiyazı olup, bu çalışma laboratuvarımızda barsak amebiyazı tanısında dışkı örneklerinde E. histolytica’ya özgü lektin antijenini saptamaya yönelik ilk çalışmadır. Bu amaçla TechLab Entamoeba histolytica II ELISA testi kullanılmıştır. Buna ek olarak, barsak amebiyazı ön tanısı alan hastaların dışkı örnekleri, Miniparasep konsantrasyon, Ecofix-Ecostain trikrom metodlarıyla mikroskopik olarak incelenmiş ve Robinson besiyerine ekimleri yapılarak amip üremesi yönünden takip edilmişlerdir. Çalışmamıza barsak amebiyazı klinik ön tanısı alan 54 hasta dahil edilmiştir. TechLab Entamoeba histolytica II antijen testiyle hastaların %5,6’sında (n: 3) E. histoloyica’ya özgü lektin antijeni tesbit edilmiştir. Ecofix-Ecostain trikrom boyama yöntemiyle 5 (%9,3) hastaya ait dışkı örneğinde amip trofozoitleri görülmüş olup, bu hastalar içinde TechLab Entamoeba histolytica II testiyle lektin antijeni saptanan 3 hasta da yer almaktadır. Miniparasep konsantrasyon ve Ecofix-Ecostain Trikrom yöntemlerinden herhangi biriyle, 11 (%20.4) hastaya ait dışkı örneklerinde amip kistleri görülmüştür. Her iki yöntemle birden amip kisti saptanan dışkı örneği sayısı 5 olup iki yöntem arasında uyum mevcuttur (r: 0.567, p:0.000). Robinson besiyerine yapılan ekimlerde, hiçbir hastaya ait dışkı örneğinde amip üremesi tesbit edilmemiştir. Yüksek duyarlılık ve özgüllüğüne ek olarak, çalışma süresince edindiğimiz laboratuvar pratiği bize, TechLab Entamoeba histolytica II testinin laboratuvar uygulamasının çok kolay olduğunu ve bu testle üretilen sonuçların, laboratuvar personelinin yorumda zorlanmasına imkan vermeyecek şekilde net olduğunu göstermiştir. TechLab Entamoeba histolytica II testiyle arasında saptanan uyum (r:0.759, p:000) ve Miniparasep yöntemiyle konsantrasyon işlemi sonrası lugolle yapılan mikroskopik incelemeye göre amip trofozoit ve kistlerinin saptanmasında uygulama kolaylığı ve ince yapısınında daha net görülmesi dikkate alındığında, Ecofix-Ecostain trikrom yönteminin barsak amebiyazı tanısında ön tarama testi olarak rutin tanı laboratuvarlarında kullanılabileceğini göstermiştir.
Intestinal amebiasis is the most common clinical form of the symptomatic E. histolytica infections. This is the first study in our laboratory investigating E. histolytica specific lectin antigen in diagnosis of intestinal amebiasis. For this purpose, TechLab Entamoeba histolytica II ELISA assay was used. Microscopic examinations of stool samples for amebic cysts and trophozoites were performed by using both Miniparasep concentration method and Ecofix-Ecostain Trichrome technique. In addition, inoculation of stool samples into Robinson’s medium were performed for culture of amebas. Fifty-four patients with prediagnosis of intestinal amebiasis were included in the study. With TechLab Entamoeba histolytica II kit, E. histolytica specific lectin antigen was detected in 3 (5.6%) patients. By using Ecofix-Ecostain Trichrome method, amebic trophozoites were detected in the stool samples of 5 (9.3%) patients including those who were positive with TechLab Entamoeba histolytica II ELISA test. Amebic cysts were detected in 11 (20.4%) patients by using of Miniparasep or Ecofix-Ecostain Trichrome techniques. In 5 patients, amebic cysts were detected in the stool samples indicated a correlation between the two methods (r: 0.567, p:0.000). In none of the stool samples, amebic growth was observed in Robinson’s media. In addition to the high sensitivity and specificity, our laboratory practice with TechLab Entamoeba histolytica II ELISA test during the study period showed us that this method is highly user friendly producing clear results which do not lead to misinterpretation by the laboratory staff. According to the correlation between TechLab Entamoeba histolytica II test (r:0.759, p:000) and the clearity and simplicity for detectection of amebic trophozoites and cysts than microscopic interpretation with lugol after Miniparasep concentration, we suggest that Ecofix-Ecostain Trichorome staining tecnique will be use a pre-scanning test for intestinal amebiasis in routine laboratories.
Intestinal amebiasis is the most common clinical form of the symptomatic E. histolytica infections. This is the first study in our laboratory investigating E. histolytica specific lectin antigen in diagnosis of intestinal amebiasis. For this purpose, TechLab Entamoeba histolytica II ELISA assay was used. Microscopic examinations of stool samples for amebic cysts and trophozoites were performed by using both Miniparasep concentration method and Ecofix-Ecostain Trichrome technique. In addition, inoculation of stool samples into Robinson’s medium were performed for culture of amebas. Fifty-four patients with prediagnosis of intestinal amebiasis were included in the study. With TechLab Entamoeba histolytica II kit, E. histolytica specific lectin antigen was detected in 3 (5.6%) patients. By using Ecofix-Ecostain Trichrome method, amebic trophozoites were detected in the stool samples of 5 (9.3%) patients including those who were positive with TechLab Entamoeba histolytica II ELISA test. Amebic cysts were detected in 11 (20.4%) patients by using of Miniparasep or Ecofix-Ecostain Trichrome techniques. In 5 patients, amebic cysts were detected in the stool samples indicated a correlation between the two methods (r: 0.567, p:0.000). In none of the stool samples, amebic growth was observed in Robinson’s media. In addition to the high sensitivity and specificity, our laboratory practice with TechLab Entamoeba histolytica II ELISA test during the study period showed us that this method is highly user friendly producing clear results which do not lead to misinterpretation by the laboratory staff. According to the correlation between TechLab Entamoeba histolytica II test (r:0.759, p:000) and the clearity and simplicity for detectection of amebic trophozoites and cysts than microscopic interpretation with lugol after Miniparasep concentration, we suggest that Ecofix-Ecostain Trichorome staining tecnique will be use a pre-scanning test for intestinal amebiasis in routine laboratories.