Tesis

Deteksi molekular Entamoeba histolytica dari sampel feses pasien kolitis di Rumah Sakit Ciptomangunkusumo, Jakarta, Indonesia = Molecular detection of Entamoeba histolytica from stool sampels of colitis patients at Ciptomangunkusumo hospital, Jakarta, Indonesia.

Kolitis masih menjadi masalah kesehatan terutama pada anak. Kolitis dapat karena bakteri ataupun parasit. Parasit yang sering diasosiasikan sebagai penyebab kolitis adalah protozoa usus seperti Entamoeba histolytica, Blastocystis sp, Cyclospora. Kolitis amoebiasis dapat menimbulkan komplikasi serius seperti fulminant necrotizing colitis, toxic megacolon, ulkus perianal, amebiasis ekstraintestinal dan stunting pada anak akibat berkurangnya penyerapan nutrisi. Selama ini diagnosis amoebiasis masih mengandalkan pemeriksaan mikroskopis dimana tidak dapat membedakan Entamoeba histolytica dari Entamoeba non patogen seperti Entamoeba dispar dan Entamoeba moshkovskii karena morfologinya yang sangat mirip. Penelitian ini bertujuan untuk mengetahui proporsi infeksi amoeba pada penderita kolitis dengan tehnik molekular / PCR. Metode: Studi ini menggunakan metode potong lintang retrospektif. Sampel feses berasal dari pasien terduga kolitis dari Rumah Sakit Cipto Mangunkusumo (RSCM), Jakarta antara tahun 2020-2022 yang dikirim untuk pemeriksaan feses parasitologi di laboratorium Parasitologi FKUI. Pemeriksaan mikroskopis terdiri dari pemeriksaan langsung dengan larutan Lugol, konsentrasi water eter, pewarnaan tahan asam, dan kultur media Jones untuk Blastocystis sp. Pemeriksaan Coproantigen Cryptosporidium/Giardia dan direct PCR 18S rRNA Entamoeba untuk deteksi E. histolytica, E. dispar, E. moshkovskii. Data sekunder berupa lama dan frekuensi diare, karakteristik feses, komorbid serta pemberian antibiotik/antiamoeba didapatkan dari rekam medik. Hasil: Sebanyak 111 pasien kolitis dari RSCM yang diperiksa feses parasitologi antara tahun 2020-2022 dan hanya 54 sampel feses yang dapat diperiksa coproantigen Cryptosporidium/Giardia dan PCR Entamoeba. Penderita laki – laki sebanyak 69,4%(77/111). Hasil pemeriksaan terhadap 54 sampel didapati proporsi protozoa sebesar 32%(17/54) dengan penyebab terbanyak oleh Blastocystis 11%, diikuti oleh Entamoeba histolytica 9%, Entamoeba moshkovskii 6%, Entamoeba dispar 4%, Cryptosporidium 4%, dan Dientamoeba fragilis 2%. Sebanyak dua (2) sampel terdapat infeksi campuran yaitu Dientamoeba fragilis – Trichuris trichiura – E. moshkovskii dan Cryptosporidium – E. histolytica. Sebagian besar yang terinfeksi Entamoeba berusia dibawah 11 tahun. Tidak ada sampel yang terdeteksi Entamoeba pada pemeriksaan secara mikroskopis, semua Entamoeba tersebut didapat dari pemeriksaan PCR. Kesimpulan: Proporsi infeksi Entamoeba pada anak dengan dugaan kolitis di RSCM tinggi dengan penyebab utama adalah Entamoeba histolytica. Sensitivitas pemeriksaan Entamoeba secara mikroskopis sangatlah rendah sehingga perlu adanya pemeriksaan lanjutan seperti PCR Entamoeba untuk mendeteksi infeksi Entamoeba pada kolitis infektif.
Kata kunci: Kolitis, Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Anak-anak


Colitis is still a health problem, especially among children. Colitis can caused by bacteria or parasites. Parasites frequently associated with colitis are the intestinal protozoa such as Entamoeba histolytica, Blastocystis sp, or Cyclospora. Colitis amoebiasis in children may cause serious complication such as fulminant necrotizing colitis, toxic megacolon, perianal ulcers, extraintestinal amebiasis and stunting in children due to reduced absorption of nutrients. So far, the diagnosis of amoebiasis still relies on microscopic examination which cannot distinguish Entamoeba histolytica from non-pathogenic Entamoeba such as Entamoeba dispar and Entamoeba moshkovskii because they have similar morphology. This study aimed to determine the proportion of amoeba infection among colitis patients using molecular technique. Methods: This study design was a retrospective cross-sectional. Stool samples collected from patients suspected with colitis from Cipto Mangunkusumo Hospital (RSCM), Jakarta and requested fecal parasitologytest at Laboratorium Parasitology FKUI between 2020-2022. The microscopic examination consisted of direct examination with Lugol's solution, water ether concentration, acid-fast stain, and Jones media culture for Blastocystis sp. Examination of coproantigen Cryptosporidium/Giardia and direct PCR 18S rRNA of Entamoeba for detection of E. histolytica, E. dispar, E. moshkovskii. Secondary data were obtained from medical records, such as duration and frequency of diarrhea, stool characteristics, comorbidities, and administration of antibiotics/antiamoeba. Results: 111 colitis patients from RSCM were examined for fecal parasitology between 2020-2022 and only 54 stool samples could be examined for coproantigen Cryptosporidium/Giardia and Entamoeba PCR. Male sufferers were 69.4% (77/111). The results of examination of 54 samples found that the proportion of protozoa as cause of colitis was 32% (17/54) with Blastocystis sp 11%, followed by Entamoeba histolytica 9%, Entamoeba moshkovskii 6%, Entamoeba dispar 4%, Cryptosporidium 4%, and Dientamoeba fragilis 2%. There were 2 samples with mixed infections, Dientamoeba fragilis - Trichuris trichiura and E. moshkovskii - Cryptosporidium - E. histolytica infections. Most of those infected with Entamoeba are under 11 years old. No samples of Entamoeba were detected on microscopic examination, all of these Entamoeba were obtained from PCR examination. Conclusion: The proportion of Entamoeba infection in children with colitis at RSCM was high with the main etiology Entamoeba histolytica. The sensitivity of microscopic examination of Entmaoeba is very low so further analyses such as Entamoeba PCR are needed to detect Entamoeba infection in infective colitis.
Keywords: Colitis, Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Children