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Abstrak :
Tuberkulosis dengan resisten ganda obat (MDR-TB) semakin meningkat dan menjadi perhatian kesehatan masyarakat di berbagai helahan dunia, terutama di negara berkembang dimana kasus ini banyak terjadi. Data dan 52 negara yang dilaporkan oleh WHO melalui glohal project on tuherculosis drug resistance surveillance menunjukkan prevalensi MDR -TB mempunyai median 1. 8 % (antara 0 sampai 18.1 %) dan 11.1 % (antara 2.9 sampai 40.8 %) untuk strain resisten pada setiap obat. Data mengenai resistensi Mycobacterium tuberculosis khususnya data MDR-TB di Indonesia masih terbatas. Metode standar untuk menguji kepekaan Mycobacterium tuberculosis seperti metode proporsi atau rasio resistensi telah banyak digunakan secara luas namun bergantung pada medium padat dan memakan waktu yang lama. Sedangkan metode BACTEC 460 memherikan hasil yang cepat namun memerlukan peralatan yang banyak dan biaya yang mahal. Pada penelitian ini kami menguji 41 isolat klinik dari pasien MDR-TB menggunakan metode DSCP. Metode DSCP menggunakan 25 sumur yang herisi medium Middlehrook 7HI0 yang mengandung obat antituberkulosis lini kedua dengan berbagai konsentrasi. Obat antituberkulosis lini . kedua : sikloserin (CYC), prothionamid (PAM), amikasin (AMK), siprofloksasin (CIF), klofazimin (CLZ), klaritromisin (CLM), rifabutin (RIB), dan ofloksasin (OFX). Hasil dan Kadar Hambat Minimum (KHM) obat dibaca antara hari ke 12 sampai 19. Hasil pengujian 41 isolat dengan metode DSCP didapatkan angka resistensi : Rifabutin (31.7 %), klaritromisin (2l.9 %), sikloserin (17.0 %), klofazimin (14.6 %), amikasin (12.1 %), prothionamid (9.7 %), siprofloksasin (9.7 %), dan ofloksasin (7.3 %) . . ? Resistensi primer MDR-TB 4 isolat (9.75 %), resistensi sekunder MDR-TB 37 isolat (90.75 %). Resistensi 1 jenis obat 6 isolat (14.2 %), resistensi 2 jenis obat 20 isolat (48.7 %), resistensi lebih dari 3 jenis obat 1 isolat (2.4 %). Metode DSCP memberikan basil yang jelas ,mudah distandarisasi, cepat dan menunjukkan KHM yang terinci. ......Multidrug-resistant tuberculosis (MDR-TB) is an increasing public health concern in many parts of the world, especially in developing countries where most cases occur. Data from 52 countries in the World Health Organizations global project on tuberculosis drug resistance surveillance shows a median prevalence of 1.8 % (range 0 to 18.1 %) for MDR-TB strains and 11.1 % (range 2.9 to 40.8 %) for strains with any drug resistance. Data on drug resistance of Mycobacterium tuberculosis especially MDR-TB in Indonesia are very limited. Standard methods for drug susceptibility testing of Mycobacterium tuberculosis, such as the proportion method or resistance ratio method, are used generally but depend on culture on solid media and therefore time-consuming. The BACTEC 460 method is faster but demands costly equipment and expensive. In this study we examined 41 clinical isolates from patients with MDR-TB by Drug Susceptibility Culture Plate (DSCP) method. DSCP use 25 wells plate filled with Middlebrook 7HIO medium containing serial dilution of second line antituberculosis drugs. Second line antituberculosis drug: Cyclose~ne (CYC), Prothionamid (PAM), Arnikacin (AMK), Ciprofloxacin (ClP), Clofazimin (CIZ), Claritromycin (CLM), Rifabutin (RIB), and Ofloxacin (OFX). The result and MIC values are read within 12 - 19 days. Result from 41 isolates that have been tested by DSCP method showed resistance to : Rifabutin, claritromycin, cycloserine, clofazimin, amikacin, prothionamid, ciprofloxacin , and ofloxacin were 31.7%, 21.9%, 17.1%, 14.6%, 12.2%, 9.7%, 9.7%, and 7.3% respectively. Primary resistance MDR-TB was 4 isolates (9.75 %) and secondary resistance MDR-TB was 37 isolates (90.75 %). Resistance to 1 drug was 6 isolates (14.2 %), resistance to 2 drugs was 20 (48.7 %) and resistance more 3 drugs was 1 (2.4 %). DSCP method potentially gives better result as it can be very well standardized, faster and provides detailed MIC (Minimal Inhibitory Concentration) values.

Abstrak :
ABSTRAK
Latar belakang : Cytomegalovirus (CMV) merupakan salah satu infeksi oportunistik pada pasien dengan sindrom immunodefisiensi (AIDS). Gejala klinis dan CT scan tidak dapat menegakkan diagnosa definitif ensefalitis CMV. Oleh karena itu diperlukan uji alternatif untuk menegakkan diagnosis infeksi CMV pada pasien HIV dengan infeksi otak. Salah satu uji yang sensitif dan spesifik adalah Real Time Polymerase Chain Reaction (rPCR). Tujuan : Mendapatkan uji deteksi molekular CMV pada pasien HIV dengan tersangka infeksi otak. Metode : Penelitian dilakukan dalam 3 tahap. Tahap 1 adalah optimasi konsentrasi primer, probe, suhu annealing, volume elusi ekstraksi DNA, dan volume cetakan. Tahap 2 adalah uji spesifisitas (reaksi silang) dan uji sensitivitas (ambang batas deteksi DNA) rPCR dan tahap 3 adalah penerapan uji rPCR yang sudah dioptimasi terhadap sampel plasma, urin, dan LCS. Hasil : Kondisi optimal uji rPCR telah diperoleh dengan konsentrasi primer dan probe 0,1 μM, dengan kondisi suhu reaksi rPCR: aktivasi enzim pada 950C selama 3 menit; 45 siklus pada 950C selama 15 detik (denaturasi) dan 560C selama 1 menit (annealing dan ekstensi). Volume elusi ekstraksi DNA yang optimal untuk ketiga jenis sampel (LCS, plasma dan urin) adalah 40 μL, dan volume cetakan rPCR untuk LCS, plasma, dan urin, masing-masing adalah 5, 4, dan 3 μL. Uji rPCR mampu mendeteksi DNA pada 50.000 jumlah kopi/mL dan tidak bereaksi silang dengan Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Candida spp, Toxoplasma gondii, EBV,HSV,dan VZV. Penerapan uji rPCR pada sampel klinis memberikan hasil negatif pada semua sampel LCS, 72,22% positif pada sampel plasma, dan 72,22% positif pada sampel urin. Kesimpulan: Telah dilakukan optimasi uji rPCR dengan minimal deteksi DNA CMV 50.000 jumlah kopi/mL dan tidak bereaksi silang dengan mikroorganisme yang berpotensi menyebabkan positif palsu (false positive).ABSTRACT
Background: Cytomegalovirus (CMV) is one of opportunistic infections in patients with Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are not typical, and CT scans can not define encephalitis CMV specifically. Therefore, it is important to apply an alternative assay for sensitive and specific detection of CMV infection in HIV patients with suspected central nervous system (CNS) infections. One of the assays is real time polymerase chain reaction (rPCR). Objective: To obtain a molecular assay for detection of CMV in HIV patients with suspect CNS infections. Methods: This study was conducted in three phases. The first is optimization of concentrations of primers, probe, annealing temperature, final elution of DNA extraction, and volume of PCR template. The second is determinations of sensitivity (minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR, and the third is application of the rPCR for clinical samples of plasma, urine, and liquor cerebrospinal (LCS). Results: The rPCR reaction showed optimal concentrations of primers and probe at 0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45 cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing and extension). Final elution of DNA extraction was 40 μL and volume of PCR templates for urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimal detection of DNA at 50,000 copies/mL and was not cross-reacted with Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), Herpes Simplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR for clinical samples showed that the rPCR yielded 72.22% positive for plasma or urine, and negative for all LCS samples. Conclusion: The rPCR has been optimized in this study with minimal DNA detection at 50,000 copies/mL and was not cross-reacted with other microorganisms that are potential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patients with Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are not typical, and CT scans can not define encephalitis CMV specifically. Therefore, it is important to apply an alternative assay for sensitive and specific detection of CMV infection in HIV patients with suspected central nervous system (CNS) infections. One of the assays is real time polymerase chain reaction (rPCR). Objective: To obtain a molecular assay for detection of CMV in HIV patients with suspect CNS infections. Methods: This study was conducted in three phases. The first is optimization of concentrations of primers, probe, annealing temperature, final elution of DNA extraction, and volume of PCR template. The second is determinations of sensitivity (minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR, and the third is application of the rPCR for clinical samples of plasma, urine, and liquor cerebrospinal (LCS). Results: The rPCR reaction showed optimal concentrations of primers and probe at 0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45 cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing and extension). Final elution of DNA extraction was 40 μL and volume of PCR templates for urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimal detection of DNA at 50,000 copies/mL and was not cross-reacted with Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), Herpes Simplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR for clinical samples showed that the rPCR yielded 72.22% positive for plasma or urine, and negative for all LCS samples. Conclusion: The rPCR has been optimized in this study with minimal DNA detection at 50,000 copies/mL and was not cross-reacted with other microorganisms that are potential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patients with Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are not typical, and CT scans can not define encephalitis CMV specifically. Therefore, it is important to apply an alternative assay for sensitive and specific detection of CMV infection in HIV patients with suspected central nervous system (CNS) infections. One of the assays is real time polymerase chain reaction (rPCR). Objective: To obtain a molecular assay for detection of CMV in HIV patients with suspect CNS infections. Methods: This study was conducted in three phases. The first is optimization of concentrations of primers, probe, annealing temperature, final elution of DNA extraction, and volume of PCR template. The second is determinations of sensitivity (minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR, and the third is application of the rPCR for clinical samples of plasma, urine, and liquor cerebrospinal (LCS). Results: The rPCR reaction showed optimal concentrations of primers and probe at 0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45 cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing and extension). Final elution of DNA extraction was 40 μL and volume of PCR templates for urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimal detection of DNA at 50,000 copies/mL and was not cross-reacted with Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), Herpes Simplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR for clinical samples showed that the rPCR yielded 72.22% positive for plasma or urine, and negative for all LCS samples. Conclusion: The rPCR has been optimized in this study with minimal DNA detection at 50,000 copies/mL and was not cross-reacted with other microorganisms that are potential to cause false positive results.;Background: Cytomegalovirus (CMV) is one of opportunistic infections in patients with Aquired Immunodeficiency Syndrome (AIDS). Clinical manifestations are not typical, and CT scans can not define encephalitis CMV specifically. Therefore, it is important to apply an alternative assay for sensitive and specific detection of CMV infection in HIV patients with suspected central nervous system (CNS) infections. One of the assays is real time polymerase chain reaction (rPCR). Objective: To obtain a molecular assay for detection of CMV in HIV patients with suspect CNS infections. Methods: This study was conducted in three phases. The first is optimization of concentrations of primers, probe, annealing temperature, final elution of DNA extraction, and volume of PCR template. The second is determinations of sensitivity (minimal detection of DNA) and specificity (cross-reaction) of the optimized rPCR, and the third is application of the rPCR for clinical samples of plasma, urine, and liquor cerebrospinal (LCS). Results: The rPCR reaction showed optimal concentrations of primers and probe at 0.1 μM, with thermal cycler: 950C for 3 min (enzyme activation), followed by 45 cycles of 950C for 15 sec (denaturation) and 560C for 1 min (annealing and extension). Final elution of DNA extraction was 40 μL and volume of PCR templates for urine, plasma, and LCS was 3, 4, and 5 μL, respectively. The rPCR had minimal detection of DNA at 50,000 copies/mL and was not cross-reacted with Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Candida spp, Toxoplasma gondii, Epstein-Bar Virus (EBV), Herpes Simplex Virus (HSV) and Varicella Zoster Virus (VZV). Application of rPCR for clinical samples showed that the rPCR yielded 72.22% positive for plasma or urine, and negative for all LCS samples. Conclusion: The rPCR has been optimized in this study with minimal DNA detection at 50,000 copies/mL and was not cross-reacted with other microorganisms that are potential to cause false positive results.

Abstrak :
Resistensi antimikroba menjadi masalah kesehatan global. Infeksi bakteri resisten dapat meningkatkan biaya perawatan kesehatan, lama perawatan di rumah sakit, morbiditas dan mortalitas baik di negara maju maupun negara berkembang. Penelitian yang menghubungkan antara infeksi oleh bakteri gram negatif resisten antibiotik dengan biaya dan lama perawatan rumah sakit belum banyak dilakukan terutama di Indonesia. Penelitian ini adalah penelitian potong lintang yang melihat perbandingan biaya perawatan dan lama rawat rumah sakit pada pasien dengan infeksi bakteri gram negatif resisten antibiotik dan peka antibiotik. Pengambilan data dilakukan secara konsekutif dengan kriteria inklusi adalah pasien yang berusis ≥18 tahun dan dirawat inap dengan hasil biakan positif terdapat isolat bakteri Gram negatif. Kriteria eksklusi adalah data psien dari laboratorium mikrobiologi yang tidak sesuai dan pasien yang tidak mendapat antibiotik. Dari 359 isolat hasil penelitian didapatkan sebanyak 221 isolat (61.6%) merupakan isolat bakteri gram negatif yang resisten antibiotik. Adapun bakteri tersebut terdiri K. pneumoniae penghasil ESBL sebanyak 97 isolat (27%), E. coli penghasil ESBL sebanyak 85 isolat (23.7%), P. aeruginosa yang resisten meropenem sebanyak 11 isolat (3.1%) dan A. baumannii resisten meropenem sebanyak 28 isolat (7.8%). Hasil perhitungan biaya perawatan pasien yang terinfeksi bakteri resisten memiliki rerata sebesar Rp 26.010.218,- sedangkan pasien yang terinfeksi bakteri peka memiliki rerata biaya perawatan sebesar Rp 18.201.234,- (p<0.05). Pasien yang terinfeksi A. baumannii resisten meropenem memiliki biaya rawat inap yang paling besar, diikuti E. coli penghasil ESBL, K. pneumoniae penghasil ESBL, dan P. aeruginosa resisten meropenem. Jumlah hari rawat pasien yang terkena infeksi bakteri adalah 14 hari, dan pasien yang terkena infeksi bakteri nonresisten adalah 9 hari (p<0.05). Hasil penelitian ini memperlihatkan bahwa infeksi bakteri Gram nehatif resisten mengakibatkan biaya perawatan dan lama rawat rumah sakit meningkat secara bermakna dibandingkan pasien dengan infeksi bakteri peka antibiotik. Pemeriksaan mikrobiologi sangat penting dilakukan, agar pasien mendapatkan antibiotik yang tepat. ......Antimicrobial resistance is a global health problems. Resistant bacterial infection increases hospital costs, length of hospital stay, morbidity and mortality in both developed and developing countries. A few research has been found linking infection with antibiotic resistant Gram-negative bacteria with the hospital costs in Indonesia. This study is a cross-sectional study, analyze the comparison of hospital costs in patients with antibiotic-resistant Gram-negative bacterial infections and antibiotic sensitive infections. The sample method is consecutive non-random sampling, with inclusion criteria were patients who were aged ≥18 years and hospitalized with Gram negative bacterial positive culture. Exclusion criteria were inappropriate patient data and patients not receiving antibiotics. From 359 isolates, 221 isolates (61.6%) were antibiotic resistant Gram negative bacteria. The bacteria consisted of 97 isolates (27%) of ESBL-producing K. pneumoniae, 85 isolates (23.7%) were ESBL-producing E. coli, 28 isolates (7.8%) were meropenem-resistant A. baumannii, and 11 isolates (3.1%) were meropenem-resistant P. aeruginosa. The average hospital cost of patients with antibiotic resistant Gram-negative bacteria was Rp. 26,010,218, whereas patients with antibiotic sensitive infection was Rp. 18,201,234, - (p<0.05). Patients with meropenem resistant A. baumannii have the highest hospital costs, followed by ESBL-producing E. coli, ESBL-producing K. pneumoniae, and meropenem-resistant P. aeruginosa. The average length of hospital stay in patients with antibiotic-resistant Gram-negative bacterial infections was 14 days, whereas patients with antibiotic sensitive infection was 9 days (p<0.05). The results showed that resistant Gram-negative bacterial infection is significantly higher hospital costs and hospital stay compared to patients with antibiotic-sensitive bacterial infections. Microbiological culture is important to do, so the patients will get the right antibiotics.