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"Cryptosporidiurn sp. adalah parasit protozoa usus intraseluler yang rnenginfeksi berbagai hewan verteblata termasuk manusia dan menyehabkan penyakit kriptosporidiosis, juga merupakan agen penycbab diare yang bersifat oportunistik. Gejaia yang berulang dan angka penularan yang tinggi akan menurunkan kualitas hidup penderita sehingga diperlukan diagnosis kriptosporidiosis yang cepat secara mikroskopis pada sediaan tinja yang diwarnai.Tesis ini bcrtujuan membandingkan metode pewamaan modiiikasi tahan asam (MTA) dan auramin fenol (AF) untuk dcteksi ookista Cryploaporidiwn sp. dari sampel tirja dengan dan tanpa konsentrasi. Sensitivitas dan spesifisitas setiap metode dari tinja yang dikonsentrasi, ditentukan dengan PCR* sebagai baku emaa Penelitian ini adalah penelitian kualitatif dengan desain cross sectional menggunakan uji diagnostik. Hasil uji skrining dan tingkat agreement dihitung Dari 130 sampel tirqa yang diperiksa, 5,4%, 10%, 10%, l9,2% dan 32,3% positif Cryprosporidium sp. dengan metode MTA tanpa konsentrasi, MTA dikonsentrasi, AF tanpa konsenuasi, AF dikonsemrasi dan PCR*. I-Iasil positif ookista Cfgptosporidium sp. lebih banyak ditemukan pada sediazm tinja yang dikonsentrasi. Hasil tidak berbeda bermakna pada perbandingan basil MTA dengan dan tanpa konsentrasi(p=0,07), sedangkan hasil berbeda bcrmakna pada AF (p=0,00). Sensitivitas MTA dan AF tinja konsentrasi adalah 30,9% dan 54,8%; spesifisitas 100% dan 97,7% dibandingkan dengan PCR*. Metode pewarnaan AF memiliki nilai sensitivitas lebih tinggi, tetapi spesifisitasnya sama dengan MTA. Metode pewamaan AF dapat digunakan scbagai altematif dari pewamaan MTA untuk deteksi ookista Cfyptosporidium sp. pada sampel tinja.

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Cryptosporzdnm sp. is intestinal protozoa parasite intracellular which infect widely vertebrata include human and cause cryptosporidiosis disease, also opportunistic agent for diarrhea. Reinfection and high transmission can decrease quality of life patient, so it needs a quick diagnostic with microscopy analysis to stain fecal smears. The objective of this study is to investigate the comparison of the modified acid fast (MAF) and auramine phenol (APh) staining method in order to detecting Cryptosporidium sp. oocysts 'nom unconcentrated and concentrated fecal sample. The sensitivity and specificity of each method from concentrated fecal sample was determined with PCR* as the gold standard.

The result of the screening test and the levels of agreement were quantified This research is qualitative interpretation with cross sectional design study which using diagnostic test. Of the |30 fecal samples that has examined, 5,4%, 10%, 10%, 19,2% and 32,3% were positive Cryptosporidium sp. by the MAF unconcentrated, MAF concentrated, APh unconoentrated, APh concentrated and PCR* method respectively. The majority of positive Cryptosporidium sp. samples were found in concentrated samples.

The results have no significant differences between MAF staining with unconcentrated and concentrated fecal sample (p=0,07), but there is a significant difference for APh staining (p=0,00). In comparison with PCR* results, the sensitivities of MAF and APh concentrated methods were 30,9% and 54,8%; the specificities were l00% and 97,7% respectively. The APh staining method apparently has more sensitivity than MAF staining method, but has the same speciticity. The APh staining method proved to be a valuable altemative to MAF staining for detection of' Cryptosporidium sp. oocysts in fecal sample."

"Latar Belakang: Metode PCR dapat digunakan untuk mengidentifikasi DNA suatu organisme. ldentifikasi gen 18S rRNA sudah hanyak dipakai untuk mempelajari sejumlah organisms eukariot seperti tanaman, hewan dan protozoa termasuk Cryptosporidium sp. Penelitian terdahulu pada anak batita di daerah kumuh dan rawan banjir di Jakarta yang dideteksi secara mikroskopis dengan pcwarnaan modifikasi tahan 838111 (MTA), didapatkan prevalensi 2,l% yang rendah dibandingkan negara berkembang lain yang keadaan lingkungan dan populasinya mirip Indonesia. Deteksi Cryptosporidium sp. secara molekular di feses dengan PCR memungkinkan diagnosis lebih akurat dan cepat. Tujuan: mengembangkan metode PCR :mink deteksi gen ISS rRNA Cryptosporidium sp. pada feses yang disimpan dalam larutan kalium bikromat 2,5% selama 1 bulan. Metode: sejumlah 188 sampel feses yang disimpan dalam larutan kalium bikromat dikonsentrasikan dengan teknik air-eter, selanjutnya dilakukan ekstraksi DNA terhadap konsentrat dan sebagian lagi dipulas dengan MTA. Amplifikasi DNA Cryptosporidium terhadap gen 18S rRNA dilakukan dengan program PCR iangsung, siklus 39 kali. Hasil: pemeriksaan dengan metode MTA konsentrasi didapatkan sembilan sampel positif (4,8%) sedangkan dengan PCR didapatkan 65 sampel positif (34,6%). Kesimpulan: Larutan kalium bikromat dapat dipakai untuk penyimpanan ookista Cryptosporidfwn sp. tanpa mempengaruhi hasil PCR maupun mikroskopis.

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Background: PCR method can be used to characterize an organism DNA. Identification of 18S rRNA gene has been widely used to study eukaryotes like plants, animals and protozoa such as Cryptosporidium sp. Previous study on Cryptosporidium sp. in children under tluee years old in a slum area in Jakarta, detected by direct modiiied acid fast (MAF) showed 2.1% prevalence, which was unexpectedly much lower than other developing country with similar enviromnent and study population. Detection of Cryptosporidium sp. by molecular technique, PCR will offer more accurate and efficient diagnosis.

Objective: develop PCR method to detect Cryptosporidium sp. 18S rRNA gene of stool from stools preserved in potassium dichromate solution for 13 months.

Methods: There were l88 stool samples which have been kept in 2.5% potassium dichromate for 13 months. These samples were concentrated by water-ether technique, extracted the DNA and stained by MAF. Amplihcation of Cnprosporidium sp. ISS rRNA gene was using direct PCR for 39 cycles.

Result: of samples with MAF has found 9 positive (4,8%) and samples with PCR has presented 65 positive (34,6%).

Conclusion: Potassium dichromate solution can be used to preserve oocysts of Cryptosporidium sp since it does not interfere the PCR and microscopic examination result."