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"ABSTRACTPenyakit Tuberkulosis (TB) merupakan salah satu ancaman kesehatan yang mematikan, Pasien TB yang tidak mendapat pengobatan tepat dapat menjadi sumber infeksi di komunitas. Metode deteksi yang efektif sangat diperlukan dalam penanganan TB. Pemeriksaan biakan dahak merupakan metode baku emas (gold standard) namun memerlukan waktu relatif lama dan mahal. Pemeriksaan mikroskopik merupakan pemeriksaan yang banyak digunakan di negara endemik TB. Namun demikian metode tersebut memiliki sensitivitas yang rendah, tidak mampu dalam menentukan kepekaan obat dan memiliki kualitas yang berbeda karena dipengaruhi oleh tingkat keterampilan petugas laboratorium. Hal tersebut diharapkan dapat diatasi dengan penggunaan pemeriksaan Tes Cepat Molekular (TCM) yang lebih cepat dibandingkan uji kepekaan dan dapat mengidentifikasi keberadaan kuman TB yang resistens terhadap rifampisin. Metode pemeriksaan TCM yang saat ini digunakan di Indonesia dengan menggunakan Xpert MTB/Rif. Penggunaan Xpert MTB/Rif telah direkomendasikan oleh WHO sejak tahun 2010. Sampai akhir 2017, telah terpasang 51 mesin TCM di Provinsi DKI Jakarta. Sehingga dilakukan penelitian untuk megetahui pemanfaatannya dengan melihat utlisasi TCM dan faktor yang mempengaruhi utilisasinya. Penelitian dilakukan dengan mengumpulkan data primer dan data sekunder. Data primer didapat dengan wawancara terhadap petugas laboratorium di 13 fasilitas kesehatan yang terdapat di Provinsi DKI Jakarta, sedangkan data sekunder didapat dari laporan utilisasi TCM tahun 2017. Data primer dianalisis untuk mendapatkan informasi hal-hal yang mungkin mempengaruhi utilisasi TCM di suatu fasilitas kesehatan. Sedangkan data sekunder dianalisis untuk mendapatkan infromasi utilisasi TCM, tipe terduga yang diperiksa dengan TCM dan hasil pemeriksaan TB dengan TCM. Dari hasil peneltian didapatkan bahwa utlisasi TCM tahun 2017 sebesar 23,28%. Faktor yang mempengaruhinya yaitu masih ada fasilitas kesehatan yang belum memiliki jejaring pemeriksaan untuk pemeriksaan TCM serta masih adanya permintaan pemeriksaan mikroskopik BTA untuk diagnosis walaupun telah tersedia alat TCM di fasilitas kesehatan tersebut. Terkait dengan hal itu, maka jejaring untuk pemeriksaan TCM harus tersedia untuk seluruh fasilitas kesehatan yang telah terpasang alat TCM serta sosialisasi kepada klinisi atau dokter peminta pemeriksaan TB mengenai teknologi pemeriksaan TCM, alur pemeriksaan dan perawatan terduga TB, permintaan dan interpretasi hasil pemeriksaan TCM penting untuk dilakukan.

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ABSTRACTTuberculosis (TB) is one of the deadliest health threats, TB patients who do not receive proper treatment can be a source of infection in the community. Effective detection methods are needed in handling TB. Sputum culture is a gold standard method but takes along time and costs are quite expensive. Microscopic examination is an examination that is widely used in endemic TB countries. However, this method has a low sensitivity, is unable to determine drug sensitivity and has different qualities because it is influenced by the level of skill of laboratory technician. This is expected to be overcome by the use of Rapid Molecular Test which is faster than sensitivity testing and can identify the presence of M. tuberculosis that are resistant to rifampicin. Until the end of 2017, 51 machines have been installed in DKI Jakarta Province. So research is conducted to find out its utilization by looking at the utilization value of Rapid Molecular Test and the factors that influence its utilization. The study was conducted by collecting primary data and secondary data. Primary data was obtained by interviewing laboratory officers in 13 health facilities in the DKI Jakarta Province, while secondary data was obtained from the 2017 TCM utilization report. Primary data were analyzed to obtain information on matters that might affect TCM utilization in a health facility. While secondary data were analyzed to obtain information on TCM utilization, the type of TB suspect examined by TCM and the results of TB testing with TCM. From the results of the research, it was found that the utilization of Rapid Molecular Test in 2017 was 21.74%. The factors that influence it are that there are still laboratory that do not have a laboratory network for Rapid Molecular examination and there is still a demand for AFB examination for diagnosis even though rapid molecular equipment is available at the laboratory. Related to this, the laboratory network for rapid molecular examinations must be available for all laboratories that have installed Rapid Molecular machine. Socialization to clinicians who requesting TB examination regarding Rapid Molecular examination technology, TB diagnostic alghorithms, requests and interpretation of Rapid Molecular examination results must be done."

"This book is a collection of the most current practices relevant to the clinical molecular laboratorian. It begins with an introductory section on techniques and procedure. It then presents four separate sections on infectious disease, oncology, pre/post-natal, and identity testing, with specific chapters clearly outlining clinical protocols used in daily practice."

"Latar belakang: Hipoksia mengakibatkan peningkatan ROS. Hingga saat ini, belum banyak diketahui mengenai peran MnSOD ? enzim antioksidan endogen utama ? pada respons adaptasi sel terhadap hipoksia. Penelitian ini bertujuan menganalisis ekspresi mRNA dan aktivitas spesifi k MnSOD pada darah, jantung dan otak tikus yang diinduksi hipoksia sistemik.Metode: 25 ekor tikus Sprague Dawley diinduksi hipoksia sistemik di dalam ruang hipoksia (8-10% O2) selama 0, 1, 7, 14 atau 21 hari. Ekspresi relatif mRNA MnSOD dianalisis menggunakan Real Time RT-PCR. Aktivitas spesifi k MnSOD diukur dengan metode inhibisi xantin oksidase.Hasil: Ekspresi relatif mRNA MnSOD pada darah dan jantung tikus menurun selama fase awal induksi hipoksia sistemik (hari ke 1) dan meningkat setelah hari ke 7, sedangkan ekspresi mRNA pada otak meningkat sejak hari ke 1 dan mencapai kadar maksimum pada hari ke 7. Hasil pengukuran aktivitas spesifi k MnSOD selama awal induksi hipoksia sistemik menyerupai hasil ekspresi mRNA. Pada kondisi hipoksia yang sangat lanjut (hari ke 21), aktivitas spesifi k MnSOD pada darah, jantung dan otak menurun secara signifi kan. Ekspresi mRNA MnSOD pada ketiga jaringan tersebut selama hari ke 0-14 induksi hipoksia sistemik berkorelasi positif dengan aktivitas spesifi knya. Selain itu, ekspresi mRNA dan aktivitas spesifi k MnSOD pada jantung berkorelasi kuat dengan hasil pada darah.Kesimpulan: Ekspresi MnSOD pada fase awal dan lanjut induksi hipoksia sistemik mengalami regulasi yang berbeda. Ekspresi MnSOD pada otak berbeda dengan pada darah dan jantung, menunjukkan bahwa jaringan otak dapat lebih bertahan pada induksi hipoksia sistemik dibandingkan jantung dan darah. Pengukuran ekspresi MnSOD di dalam darah dapat digunakan untuk menggambarkan ekspresinya di dalam jantung pada keadaan hipoksia sistemik.

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AbstractBackground: Hypoxia results in an increased generation of ROS. Until now, little is known about the role of MnSOD - a major endogenous antioxidant enzyme - on the cell adaptation response against hypoxia. The aim of this study was to determine the MnSOD mRNA expression and levels of specifi c activity in blood, heart and brain of rats during induced systemic hypoxia.Methods: Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia in an hypoxic chamber (at 8-10% O2) for 0, 1, 7, 14 and 21 days, respectively. The mRNA relative expression of MnSOD was analyzed using Real Time RT-PCR. MnSOD specifi c activity was determined using xanthine oxidase inhibition assay.Results: The MnSOD mRNA relative expression in rat blood and heart was decreased during early induced systemic hypoxia (day 1) and increased as hypoxia continued, whereas the mRNA expression in brain was increased since day 1 and reached its maximum level at day 7. The result of MnSOD specifi c activity during early systemic hypoxia was similar to the mRNA expression. Under very late hypoxic condition (day 21), MnSOD specifi c activity in blood, heart and brain was signifi cantly decreased. We demonstrate a positive correlation between MnSOD mRNA expression and specifi c activity in these 3 tissues during day 0-14 of induced systemic hypoxia. Furthermore, mRNA expression and specifi c activity levels in heart strongly correlate with those in blood.Conclusion: The MnSOD expression at early and late phases of induced systemic hypoxia is distinctly regulated. The MnSOD expression in brain differs from that in blood and heart revealing that brain tissue can possibly survive better from induced systemic hypoxia than heart and blood. The determination of MnSOD expression in blood can be used to describe its expression in heart under systemic hypoxic condition. "

"This volume brings together contributions from experts in the field of Pasteurella research. Its covers areas such as comparative genomics, pathogenic mechanisms, bacterial proteomics, as well as a detailed description and analysis of PMT and its interaction with host tissues, cells, immune system, and signalling pathways."

"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."

"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."