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"ABSTRAKCandida krusei merupakan lima besar spesies Candida terbanyak penyebab kandidosis. Candida krusei sulit diobati karena memiliki resistensi primer terhadap flukonazol. Data pola kepekaan C. krusei di Jakarta berbeda dengan di dunia karena masih ditemukan isolat sensitif terhadap flukonazol. Tujuan penelitian ialah mengidentifikasi isolat C. krusei isolat Jakarta yang peka dan resisten terhadap flukonazol. Sampel penelitian menggunakan enam isolat uji C. krusei (empat resisten dan dua sensitif flukonazol) dan dua isolat kontrol kualitas yaitu C. albicans dan C. parapsilosis yang merupakan isolat koleksi Laboratorium Mikologi Departemen Parasitologi Fakultas Kedokteran Universitas Indonesia. Analisis yang dilakukan adalah fenotipik berupa makroskopik, mikroskopik serta biokimia. Analisis molekular menggunakan teknik PCR pada regio ITS (NCBI dan ISHAM) dan D1/D2 (NCBI). Hasil perbandingan sekuens ITS antar isolat uji didapatkan satu perbedaan basa pada urutan basa 465. Hasil perbandingan sekuens regio D1/D2 tidak ditemukan perbedaan basa. Analisis multi gene sequence menunjukkan kemiripan dengan isolat C. krusei CBS 5147 yang sensitif terhadap flukonazol. Analisis filogenetik regio ITS dan D1/D2 memperlihatkan isolat uji merupakan C. krusei. Identifikasi ke enam isolat Jakarta berdasarkan analisis fenotipik dan analisis molecular menunjukkan bahwa semuanya merupakan C. krusei.

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ABSTRACTCandida krusei is one of the big five species of Candida caused disease. Candida krusei becomes difficult to treat due to the primary resistance against fluconazole. Examination data of resistance in C. krusei in Jakarta contrast to the world is isolates which are still sensitive to fluconazole. The objective of this study was identified of C. krusei of Jakarta's isolates which is sensitive and resistant fluconazole. The samples used 6 isolates C. krusei (four resistant and two sensitive fluconazole) and two isolates quality control the C. albicans and C. parapsilosis were cultured collection of Mycology Laboratory Parasitology's Department, Faculty of Medicine, Universitas Indonesia. Analysis phenotypic using by macroscopic, microscopic and biochemistry. Analysis molecular was done using PCR method for ITS (NCBI and ISHAM) and D1/D2 (NCBI) regions. The results of ITS sequences comparison with isolates test were found one difference base at 465. The results of D1/D2 sequence comparison did not find the difference of sequences base, and analysis of multi Gene sequence showed the isolated test had a similarity with C. krusei CBS 5147 which sensitive fluconazole. Phylogeny of the region ITS and D1/D2 showed the isolates test was C. krusei. Identification six isolates based on analysis phenotypes and analysis of molecular was C. krusei."

"ABSTRAKThis study aimed to investigate the prevalence of the pfdhps and pfdhfr polymorphisms in southern Somalia. The genetic polymorphisms of both genes were analyzed by nested PCR-RFLP. A total of 150 samples were collected; of these, 101 were shown to be positive for Plasmodium (96 P. falciparum and 5 P. vivax) by nested PCR, the remaining 49 were PCR negative. Of the 96 Plasmodium falciparum isolates, 88 were successfully amplified for pfdhps and pfdhfr polymorphisms. The mutations occurring in the pyrimethamine resistance gene (pfdhfr) at codons 51, 59 and 108 were 59 (67.0%), 51 (58.0%) and 83 (94.3%) isolates, respectively. Sulfadoxine resistance-associated mutations in the pfdhps gene at codons 437, 540 and 581 were found in 41 (46.6%), 43 (48.9%) and 13 (14.8%) samples, respectively. The analysis of pfdhfr and pfdhps combination revealed that 27 (30.7%) isolates harbor the quintuple mutations (I51 R59 N108 - G437 E540 A581 and I51 R59 N108 - G437K540G581). The prevalence of single mutation, triple mutations, quadruple mutations and double mutations haplotypes were 19.3%, 18.2%, 15.9% and 12.5%, respectively. Additionally, sextuple mutations were observed at 2 isolates (2.3%). This study shows that the pfdhfr/pfdhps mutant alleles have moderately declined compared to a previous study, but still remain high. "

"Kendala utama dalam diagnosis penyakit tuberkulosis (TB) paru di Indonesia adalah sulitnya pengeluaran sputum sebagai spesimen diagnostik dari pengidap TB paru, terutama pada kelompok anak dan lansia. Pengembangan spesimen alternatif, seperti urine, sangat dibutuhkan untuk meningkatkan notifikasi kasus TB paru pada subjek yang belum terdiagnosis secara optimal. Penelitian ini mengevaluasi potensi urine pada 60 sampel terkonfirmasi TB paru positif, sebagai studi kasus-kontrol dengan subjek yang memiliki kondisi target. Tujuan penelitian ini meliputi evaluasi multiplex polymerase chain reaction (PCR), untuk mendeteksi gen ESAT6, IS6110, dan MPT64, serta real-time PCR untuk deteksi gen ESAT6 dalam menunjang diagnosis TB paru. Selain itu, penelitian ini diharapkan dapat menggambarkan nilai sensitivitas masing-masing metode dan gen diagnostik yang digunakan. Metode penelitian meliputi preparasi sampel urine, isolasi DNA, kuantifikasi DNA, amplifikasi DNA (multiplex PCR dan real-time PCR), elektroforesis DNA, dan analisis data. Hasil evaluasi menunjukkan kemurnian total DNA yang diisolasi dari urine berdasarkan A260/A280   (Mean 3,08  SD 1,08). Evaluasi multiplex PCR dengan gen deteksi ESAT6, IS6110, dan MPT64 memberikan nilai sensitivitas sebesar 68,3% (41/60), dan real-time PCR dengan gen deteksi ESAT6 sebesar 71,67% (43/60). Nilai sensitivitas real-time PCR diperoleh dengan ketentuan limit of detection (LOD) sebesar 13,04 kopi/μl. Nilai sensitivitas kedua metode tersebut menunjukkan bahwa urine dapat menjadi spesimen alternatif untuk pendeteksian Mycobacterium tuberculosis (Mtb) secara molekuler.

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The main obstacle in the diagnosis of pulmonary tuberculosis (TB) in Indonesia is the difficulty of extracting sputum, as a diagnostic specimen for people with pulmonary TB, especially in the group of children and the elderly. The development of alternative specimens, such as urine, is urgently needed to increase the notification of pulmonary TB cases in subjects who have not been diagnosed optimally. This study evaluated the urine potency of 60 samples of confirmed positive pulmonary TB, as a case-control study with subjects with the target condition. The objectives of this study include the evaluation of multiplex polymerase chain reaction (PCR), to detect the ESAT6, IS6110, and MPT64 genes, as well as real-time PCR to detect the ESAT6 gene in supporting the diagnosis of pulmonary TB. In addition, this study is expected to describe the sensitivity value of each diagnostic method and gene used. Research methods include urine sample preparation, DNA isolation, DNA quantification, DNA amplification (multiplex PCR and real-time PCR), DNA electrophoresis, and data analysis. The evaluation results showed the total purity of DNA isolated from urine based on A260/A280   (Mean 3,08  SD 1.08). Evaluation of multiplex PCR with ESAT6, IS6110, and MPT64 detection genes gave a sensitivity value of 68,3% (41/60), and real-time PCR with ESAT6 detection genes of 71,67% (43/60). Real-time PCR sensitivity value was obtained with the limit of detection (LOD) of 13,04 copies/μl. The sensitivity values of both methods indicate that urine can be an alternative specimen for molecular detection of Mycobacterium tuberculosis."