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"Clostridium difficile is the most important cause of antibiotic associated diarrhea, and pseudomembranous colitis, a severe infection of the colon. Strain Clostridium difficile produce two potent toxin, toxin A (enterotoxin) and toxin B (cytotoxin). These two toxins are both responsible for the diarrhea and inflammation seen in patients treated due to infection, especially the broad spectrum antibiotics. Direct detection of Clostridium difficult cytotoxin front faecal specimen using mammalion tissue culture lines is considered the standard diagnostics test of Clostridium difficult infection. This test is very sensitive but requires a minimum two days to complete. In order to improve the threshold of diagnosis and treatment, a number of enzyme immunoassay methods have been used, with a reported sensitivity to either toxin A or toxin B."

"Latar belakang. Berbagai studi sebelumnya menunjukkan bahwa insidens kolonisasi dan infeksi C.difficile semakin meningkat, terutama pada pasien rawat inap yang mendapat terapi antibiotika. Namun belum ada penelitian yang mendapatkan data kedua insidens tersebut di Indonesia, terutama di RSCM. Tujuan. Untuk mengetahui insidens kolonisasi dan infeksi C.difficile pasien rawat inap yang mendapat terapi antibiotika di RSCM. Metode. Dilakukan studi kohort prospektif berbasis surveilans pada 96 pasien rawat inap yang mendapat terapi antibiotika di RSCM pada periode penelitian. Dilakukan pemeriksaan feses dengan uji kromatografi cepat C.DIFF QUIK CHEK COMPLETETM pada awal dan akhir penelitian. Dilakukan follow-up selama 5-7 hari perawatan pada semua pasien. Insidens kolonisasi strain non-toksigenik adalah pasien yang memiliki hasil pemeriksaan fesesnya konversi GDH/Toksin -/- saat awal perawatan menjadi GDH/Toksin +/-. Insidens kolonisasi strain toksigenik adalah pasien yang memiliki konversi GDH/Toksin -/- saat awal perawatan menjadi GDH/Toksin +/+. Insidens infeksi adalah pasien yang memiliki konversi GDH/Toksin -/- saat awal perawatan menjadi GDH/Toksin +/+ yang disertai satu atau lebih gejala yang berhubungan dengan infeksi C.difficile. Hasil. Dari 96 subjek penelitian, 13 subjek mengalami kolonisasi non-toksigenik; 8 subjek mengalami kolonisasi toksigenik; 9 subjek mengalami infeksi. Terdapat 11 subjek yang mengalami gejala klinis, namun hasil pemeriksaan fesesnya tidak ditemukan toksin yang positif (2 subjek hanya mengalami kolinisasi non-toksigenik dan 9 subjek tidak mengalami kolonisasi atau infeksi) sehingga dianggap bukan merupakan infeksi C.difficile. Kesimpulan. Insidens kolonisasi C.difficile adalah 22%, dimana kolonisasi strain non-toksigenik adalah 14% (IK95% 13-16) dan strain toksi.

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Background. Previous studies showed that there have been a significant increasing of the incidence of C.difficile colonization and infection, particularly among hospital inpatients prescribed antibiotics. However, there is no such data available in Indonesia, mainly at Cipto Mangunkusumo Hospital.

Objective. To determine the incidence of Clostridium difficile colonization and infection among hospital inpatients prescribed antibiotics at Cipto Mangunkusumo Hospital.

Methods. A surveillance-based prospective cohort study was conducted on 96 inpatients prescribed antibiotics at Cipto Mangunkusumo Hospital during the study period. All patient was followed-up for 5-7 days hospitalization. We obtained rectal swabs or stool samples on admission and day 5-7 of hospitalization and performed a rapid chromatography test C.DIFF QUIK CHEK COMPLETETM to determine colonization or infection. Incidence of non-toxigenic colonization was defined as a conversion of baseline result GDH/toxin -/- into GDH/toxin +/- as the second result. Incidence of toxigenic colonization was defined as as a conversion of baseline result GDH/toxin -/- into GDH/toxin +/+ as the second result. Incidence of infection was defined as a conversion of baseline result GDH/toxin -/- into GDH/toxin +/+ as the second result, accompanied by one or more C.difficile infection-associated clinical symptoms.

Results. A total of 96 subjects were included in the study; 13, 8 and 9 had a non-toxigenic colonization, toxigenic colonization, and infection, respectively. 11 subjects with clinical symptoms could not be determined whether they had a C.difficile infection because of the “toxin-negative” findings from their stool examination (2 subjects had non-toxigenic colonization and 9 subjects had neither colonization nor infection).

Conclusion. The incidence of C.difficile colonization was 22%, which 14% (95% CI 13-16) was the incidence of non-toxigenic colonization and 8% (95% CI 7-10) was the incidence of toxigenic colonization. The incidence of C.difficile infection was 9% (95% CI 8-11)."

"Infeksi Clostridium difficile toksigenik meningkat tajam pada satu dekade terakhir, menyebabkan pseudo membran colitis (PMC) dan Clostridium difficile associated diarrhea (CDAD). Salah satu faktor risikonya adalah penggunaan antibiotik. Tujuan penelitian adalah mengetahui prevalensi dan gambaran karakteristik subyek dengan Clostridium difficile toksigenik serta menilai kemampuan rapid test toksin terhadap real time PCR. Subyek penelitian prospektif ini adalah 90 subyek dewasa dengan terapi antibiotik lebih dari 2 minggu. Hasil pemeriksaan menggunakan rapid test dan real time PCR disajikan dalam tabel 2x2, dilakukan uji statistik dengan chi square. Hasil penelitian menunjukkan 2 spesimen dieksklusi karena hasil invalid, 24 spesimen positif dan 64 negatif dengan rapid test toksin; 33 spesimen positif dan 55 negatif dengan real time PCR. Prevalensi Clostridium difficile toksigenik berdasar rapid test toksin adalah 27,3% dan real time PCR 37,5%. Terdapat perbedaan bermakna antara konsistensi feses dan jumlah antibiotik dengan terdeteksinya Clostridium difficile toksigenik (p<0,05). Terdapat hubungan antara lama terapi antibiotik dengan terdeteksinya Clostridium difficile toksigenik menggunakan real time PCR (p=0,010, RR=2,116). Sensitivitas, spesifisitas, nilai duga positif, nilai duga negatif, rasio kemungkinan positif dan rasio kemungkinan negatif rapid test toksin terhadap real time PCR berturut-turut adalah 69,7%; 98,2%; 95,8%; 84,4%; 39,2 dan 0,31. Dari hasil penelitian disimpulkan bahwa prevalensi Clostridium difficile di RSCM lebih tinggi dibanding Malaysia, Thailand dan India; subyek dengan terapi antibiotik lebih dari 4 minggu berisiko terdeteksi Clostridium difficile toksigenik 2 kali lebih besar dibanding subyek dengan terapi antibiotik kurang dari 4 minggu; rapid test toksin dapat digunakan sebagai alat deteksi Clostridium difficile toksigenik.

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Toxigenic Clostridium difficile infection have increased sharply in the last decade, causing a pseudo membrane colitis (PMC) and Clostridium difficile associated diarrhea (CDAD). One of the biggest risk factor is the use of antibiotics. The purpose of the study was to determine the prevalence and characteristics of subjects with toxigenic Clostridium difficile and assess the ability of the toxin rapid test compared to real-time PCR. Ninety adult subjects with antibiotic therapy more than 2 weeks were enrolled to this prospective study. The results of toxin rapid test and real-time PCR were presented in 2x2 table, statistical tests was calculated with chi square. Two specimens were excluded due to invalid results. The results showed 24 positive and 64 negative specimens by toxin rapid test; 33 positive and 55 negative specimens by real-time PCR. The prevalence of toxigenic Clostridium difficile based on toxin rapid test were 27.3% and 37.5% by real-time PCR. There were significant differences between stool consistency and number of antibiotics that were used with the detection of toxigenic Clostridium difficile. There was a relationship between duration of antibiotic therapy with detection of toxigenic Clostridium difficile using real-time PCR (p = 0.010, RR = 2.116). Sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio of toxin rapid test against real-time PCR were 69.7%; 98.2%; 95.8%; 84.4%; 39.2 and 0.31, respectively. The study concluded that the prevalence of Clostridium difficile in RSCM was higher than Malaysia, Thailand and India; subjects with antibiotic therapy for more than 4 weeks had double risk to have toxigenic Clostridium difficile than subjects with antibiotic therapy for less than 4 weeks and toxin rapid test could be used as a tool to detect toxigenic Clostridium difficile."

"Latar belakang: Clostridium difficile merupakan bakteri anaerob gram positif yang sering menyebabkan diare pada pasien yang dirawat di rumah sakit. Manifestasi klinis diare karena C. difficile bervariasi dapat berupa diare ringan sampai keadaan klinis yang berat seperti kolitis, komplikasi megakolon toksik, perforasi, serta syok. Faktor risiko yang berperan meningkatkan diare karena C. difficile salah satunya adalah pengunaan antibiotik namun masih dapat disebabkan oleh hal lainnya. Penelitian ini merupakan studi untuk mengetahui prevalens, faktor risiko, dan gambaran klinis diare karena C. difficile pada anak. Metode: Penelitian ini merukapan studio potong lintang, dilakukan pada pasien 105 anak dengan keluhan diare di Poliklinik Anak dan ruang rawat inap pada bulan Mei 2019 sampai Januari 2020 dengan mendeteksi antigen toksin A/B C. difficile menggunakan metode ELISA. Hasil: Prevalens diare pada anak karena C. difficile sebesar 13,3%.Usia kurang dari 2 tahun meningkatkan risiko kejadian diare karena C. difficile 3,84 kali dibandingkan dengan pasien usia lebih dari 2 tahun dan penggunaan PPI atau H2 antagonis meningkatkan risiko terjadinya diare karena C. difficile 5,48 kali dibandingan dengan kelompok yang tidak menggunakan PPI atau H2 antagonis. Semua subyek menderita diare karena C. difficile memiliki riwayat penggunaan antibiotik. Golongan sefalosporin merupakan antibiotik yang dominan terkait dengan diare karena C. difficile (92,9%), diikuti aminoglikosida 7,1%. Gambaran klinis pasien diare karena C. difficile pada penelitian ini sebagian besar mengalami frekuensi diare 6-9 kali/24 jam, lama diare 14 hari,  nyeri perut, diare dengan dehridrasi berat ataupun ringan, leukosit tinja 10/LPB, dan terdapat darah samar tinja. Diagnosis penyakit yang mendasari pada penelitian ini meliputi infeksi paru 4 subyek, penyakit lain (kongenital dan malnutrisi) 4 subyek,  penyakit hematologi dan onkologi 3 subyek, penyakit imunologi 2 subyek, dan neurologi 1 subyek. Kesimpulan: Penggunaan PPI atau H2 antagonis serta usia kurang dari 2 tahun meningkatkan risiko kejadian diare karena C. difficile. Semua subyek yang mengalami diare karena C. difficile memiliki riwayat penggunaan antibiotik lebih dari tujuh hari.

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Background and aim: Clostridium difficile is a gram-positive anaerobic bacterium that often causes diarrhea in patients who are hospitalized. Clinical manifestations of diarrhea due to C. difficile can vary from mild diarrhea to severe clinical conditions such as colitis, toxic megacolon complications, perforation, and shock. Risk factors that play a role in increasing diarrhea due to C. difficile one of which is the use of antibiotics but can still be caused by other things. This study is a study to determine the prevalence, risk factors, and clinical picture of diarrhea due to C. difficile in children. Methods: This study was a cross-sectional study, conducted on 105 pediatric patients with diarrhea complaints in the Children's Polyclinic and inpatients in May 2019 to January 2020 by detecting C. difficile A/B toxin antigen using the ELISA method. Results: The prevalence of diarrhea in children due to C. difficile is 13.3%. Age less than 2 years increased the risk of occurrence of diarrhea due to C. difficile 3,84 times compared with patients aged more than 2 years and the use of PPI or H2 antagonists increased the risk of diarrhea due to C. difficile 5,48 times compared to the group who did not use PPI or H2 antagonists. All subjects suffered from diarrhea due to C. difficile had a history of antibiotic use. Cephalosporins are the dominant antibiotics associated with diarrhea due to C. difficile (92.9%), followed by aminoglycosides 7.1%. The clinical features of diarrhea patients due to C. difficile in this study are the frequency of diarrhea 6-9 times/24 hours, duration of diarrhea 14 days, abdominal pain, diarrhea with severe or mild dehridration, stool leukocytes 10/LPB, and fecal faint blood. Diagnosis of the underlying disease in this study included 4 subjects lung infection, other diseases (congenital and malnutrition) 4 subjects, hematological and oncological diseases 3 subjects, immunological diseases 2 subjects, and neurology 1 subject.

Conclusion: The use of PPI or H2 antagonists and age less than 2 years increases the risk of diarrhea due to C. difficile. All subjects who had diarrhea due to C. difficile had a history of antibiotic use for more than seven days.

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"Ruang lingkup dan cara penelitian: Clostridium docile merupakan salah satu kuman anaerob penyebab infeksi nosokomial. Infeksinya dapat berupa Antibiotic Associated Diarrheae (AM) ataupun Pseudomembranous Colitis (PMC). Bayi dan neonates dianggap sebagai sumber penyebab infeksi nosokomial oleh kuman tersebut karena organisme ini ditemukan sebagai flora normal dalam saluran pencernaannya. Penelitian ini dilakukan untuk mengetahui persentase anak berumur kurang dari 1 tahun yang dalam fesesnya ditemukan Clostridium difficile toksigenik. Deteksi toksin dilakukan dengan uji koaglutinasi lateks terhadap filtrat kultur organisme dan dibandingkan dengan uji sitotoksisitas menggunakan biakan sel BHK-21. Isolasi organisme dilakukan pada media Cycloserine Cefoxitin Fructose Agar (CCFA) dan Cycloserine Cefoxitin Manitol Agar (CCMA). Penggunaan media CCMA bertujuan untuk memperoleh cara identifikasi Clostridium dicile yang mudah dan praktis. Hasil dan kesimpulan : Ditemukan 11,7 % anak berumur kurang dari 1 tahun mengandung Clostridium difficile toksigenik dalam fesesnya. Uji koaglutinasi lateks yang dilakukan terhadap filtrat kultur organisme memberikan nilai sensitivitas sebesar 100 %, nilai spesifisitas 100 %, nilai prediktif positif 100 % ,don nilai predlktif negatif 100 % ,dengan menggunakan uji sitotoksisitas sebagai gold standard. Jumlah dan jenis bakteri yang tumbuh pada media CCFA dan CCMA adalah sama, koloni Clostridium dif,ficile yang tumbuh pada media CCMA memiliki warna yang spesifik yaitu loaning sedang koloni spesies Clostridium lain berwarna merah muda. Dari hasil ini dapat disimpulkan bahwa 11,7 % anak membawa Clostridium difficile toksigenik dalam saluran cernanya, uji koaglutinasi lateks dengan filtrat kultur organism memberikan hasil yang sama dengan uji sitotoksisitas dengan biakan sel BHK-21, penggunaan media CCMA tidak berpengaruh terhadap tingkat isolasi Clostridium difficile bahkan memberikan gambaran koloni yang lebih spesifik sehingga mempermudah dan mempersingkat waktu identifikasi."

"Latar Belakang: Virus Newcastle Disease (ND) menyebabkan kerugian ekonomi yang sangat tinggi pada peternakan unggas. Walaupun penggunaan vaksinasi merupakan pilihan terbaik saat ini dalam mencegah infeksi virus, namun dalam suatu kondisi dibutuhkan pengembangan antiviral. Hingga saat ini langkah terapi profilaktik terhadap infeksi virus pada peternakan unggas belum pernah dikembangkan, sedangkan penggunaan antiviral yang beredar saat ini sangat tidak mungkin digunakan dengan pertimbangan ekonomi. Penelitian ini bertujuan untuk mengembangkan suatu senyawa berupa sialidase asal bakteri Clostridium perfringens yang berpotensi sebagai antiviral dan membuktikan efek penghambatan infeksi virus sehingga dapat digunakan sebagai terapi profilaktik terhadap infeksi virus ND.Metode: Penelitian diawali dengan memproduksi enzim sialidase yang berasal dari hasil supernatan kultur bakteri C. perfringens tipe A. Sialidase dipurifikasi dengan metode presipitasi ammonium sulfat, ion exchange chromatography dan affinity chromatography. Sialidase tersebut selanjutnya diuji aktivitas dan stabilitasnya terhadap beberapa pH dalam waktu inkubasi tertentu. Uji kemampuan hidrolisis reseptor sialic acid dan toksisitas terhadap sel dilakukan pada beberapa dosis pemberian sialidase menggunakan sel kultur primer Chicken Embryonic Fibroblast (CEF). Pembuktian kemampuan sialidase dalam menghambat infeksi virus pada sel host dilakukan dengan menghitung viral copy number dan ekspresi gen penyandi molekul sitokin yang berperan terhadap respon sel akibat infeksi virus ND.Hasil: Sialidase dari bakteri C. perfringens tipe A pada penelitian ini mampu diproduksi dengan efisien secara native dan dapat dimurnikan sehingga diperoleh aktivitas spesifik sebesar 75 U/mg serta stabil selama 72 jam pada suhu 37℃. Dosis tertinggi sialidase yang dapat ditolerir oleh sel CEF yakni sebesar 187,5 mU/ml dan mampu menghidrolisis reseptor sialic acid pada permukaan sel. Pemberian sialidase pada dosis tertinggi hingga dosis terendah mampu menurunkan secara drastis replikasi virus pada sel host. Hal tersebut juga didukung berdasarkan pengamatan terhadap perbedaan ekspresi gen penyandi molekul sitokin pada sel yang ditreatment dengan sialidase dibandingkan kontrol infeksi virus NDKesimpulan: Sialidase asal bakteri C. perfringens tipe A berpotensi dapat digunakan sebagai terapi profilaksis antivirus melalui aktivitas competitive inhibition terhadap reseptor sialic acid pada sel host.

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Introduction: Newcastle Disease (ND) virus causes very high economic losses on poultry farms. Although vaccination is the best option in preventing viral infections, but under certain conditions development of antivirals is required. Prophylactic treatment against viral infection in poultry have not been developed, while the use of currently circulating antivirals is very unlikely to be used due to economic considerations. This study aims to produce a substance called sialidase from Clostridium perfringens that potential as an antiviral and demonstrate its inhibitory effect on viral infection so that it can be used as prophylactic therapy against ND virus infection.

Methods: This research was initiated by producing sialidase enzyme derived from the supernatan culture of C. perfringens type A bacteria. Sialidase was purified by ammonium sulfate precipitation method, ion exchange chromatography and affinity chromatography. The sialidase was tested for its activity and stability on several pH within a certain incubation time. Hydrolysis ability of sialic acid receptors and cell toxicity were carried out at several doses of sialidase administration using primary cultured Chicken Embryonic Fibroblast (CEF) cells. The capacity of sialidase to prevent viral infection in host cells was demonstrated by estimating the viral copy number and expression of genes encoding cytokine molecules that play a role in cell response due to ND virus infection.

Results: In this study, C. perfringens type A bacteria were able to produce sialidase then natively purified to obtain specific activity of 75 U/mg and stable for 72 hours at 37℃. The highest dose of sialidase that CEF cells can tolerate and capable to hydrolyze sialic acid receptors on the cell surface is 187.5 mU/ml. Sialidase dosages ranging from 750 mU/ml to 46.87 mU can drastically reduce viral replication in CEF cells. This is also supported by observations expression alteration of genes encoding cytokine molecules in sialidase treated cells and ND virus infection.

Conclusion: Sialidase from Clostridium perfringens tipe A has the potential to be used as prophylactic antiviral therapy through its competitive inhibition activity against sialic acid receptors on host cells."

"Human Immunodeficiency Virus (HIV) causes damage to the human immune system and the disease known as Acquired Immune Deficiency Syndrome (AIDS). This virus is a member of the Lentivirus group of viruses of the Retrovirus subfamily, which has a reverse tran-scriptase enzyme. HIV infects cells which expres CD4, mediated by gp 120. HIV infection changes the lymphocyte migration pattern, the activity of cytotoxic T cells and CD4 T cell count. The T cell CD4+ count is related to the progressivity of the disease.Anti gp 120 is the antibody most abundantly produced during HIV infection. Spesific antibody concentration for the antigens vary among individuals and single individual at different stages of the infection. Expression of the HIV antigen and/or antibody can be used to establishing the diagnosis and determine the stage of the disease. CD4+ cells count can be used to determine the stage of HIV infection, to predict the occurance of opportunistic infection and other complications, and to determine as well as to monitor therapy"