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"Beberapa jenis penyakit membutuhkan penatalaksanaan secara cepat dan tepat guna menurunkan risiko fatal penderita, salah satu diantaranya adalah difteri. Waktu sangat berharga untuk bisa menolong penderita karena keterlambatan penatalaksanaan bisa meningkatkan kematian kasus hingga 20 kali lipat. Di sisi lain, metode diagnostik konvensional sebagai gold standard membutuhkan waktu 3?5 hari sehingga diperlukan metode diagnostik alternatif untuk membantu menegakkan diagnosis difteri. Direct polymerase chain reaction (PCR) dapat menjawab tantangan atas besarnya biaya dan lamanya waktu yang dibutuhkan untuk pemeriksaan laboratorium. Penelitian bertujuan untuk mengembangkan metode direct PCR sebagi metode alternatif diagnostik difteri secara cepat, mudah dan hemat. Sebanyak 15 sampel yang terdiri dari 10 isolat Corynebacterium diphtheriae toksigenik dan 3 Corynebacterium non-diphtheriae nontoksigenik ditambah dengan 2 spesimen klinis (usap tenggorok) digunakan untuk optimasi metode direct PCR dibandingkan dengan PCR standard sebagai kontrol. Hasil penelitian menunjukkan bahwa direct PCR dapat digunakan untuk deteksi dan amplifikasi gen target dengan benar seperti halnya PCR standard sehingga pada seluruh sampel C. diphtheriae tampak pita pada 168 bp (penanda gen dtxR) dan 551 bp (penanda gen tox). Sebaliknya pada sampel lain tidak tampak pita pada kedua tempat tersebut. Direct PCR dapat mendeteksi sel bakteri sampai dengan 71 CFU/uL. Oleh karena itu, disimpulkan bahwa direct PCR dapat digunakan sebagai metode diagnostik alternatif untuk membantu menegakkan diagnosis difteri secara cepat, mudah dan hemat.

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Direct PCR: Alternative Diagnostic Method for Diagnosis of Diphtheria Rapidly, Easily and Cost Effective. Some diseases require immediate and appropriate treatment to decrease the fatality risk patients incident, for example diphtheria. Time to help patients is very crucial since delay of therapy may increase the mortality cases up to 20 times. In other hands, conventional diagnostic methods (the gold standard) for diagnosis of diphtheria is time consuming and laborious. Therefore, an alternative diagnostic method which is rapid, easy and inexpensive is needed. In this case, direct PCR has been proved to reduce time and cost in laboratory examination. This study aimed to develop direct PCR as alternative diagnostic method for diagnosis of diphtheria rapidly, easily, and inexpensive. Fifteen samples include 10 isolates of Corynebacterium diphtheriae (toxigenic) and 3 isolates of Corynebacterium non- diphtheriae (nontoxigenic) and 2 clinical specimens (throat swab) was examined by performing direct PCR method and a standard PCR method was used for optimizing the protocols. Result showed that direct PCR can be used to amplify target genes correctly as well as standard PCR. All of C. diphtheriae samples showed bands at 168 bp (dtxR gene marker) and 551 bp (tox gene marker) while no band appeared in others. Direct PCR detected at least 71 CFU/uL of bacterial cells in samples. We concluded that direct PCR can be used for alternative diagnostic method for diagnosis of diphtheria which is rapid, easy and cost effective."

"Buku ini berisi tentang gambaran uji coba PCR multipleks yang dikembangkan oleh peneliti di Badan Penelitian dan Pengembangan Kesehatan untuk identifikasi penyebab difteri secara cepat dan akurat. Hal ini penting disampaikan karena sampai dengan saat ini pemeriksaan laboratorium masih menjadi salah satu kendala dalam penatalaksanaan kasus dan pengendalian KLB difteri, khususnya di Indonesia. Jarangnya kasus difteri berimplikasi pada terbatasnya laboratorium klinik yang siap melakukan pemeriksaan sampel secara rutin. Selain itu, lamanya mendapatkan hasil pemeriksaan menggunakan metode konvensional sebagai gold standard seringkali menjadi kendala di lapangan. Oleh karena itu, buku ini mencoba menyajikan beberapa keunggulan dari pemeriksaan laboratorium difteri menggunakan teknik PCR multipleks dibandingkan dengan metode lain. Inti dari buku ini disajikan pada Bab IV dan V tentang hasil penelitian dan pembahasan. Sebagai tambahan, konsep umum atau tinjauan pustaka disajikan pada Bab II untuk memudahkan para pembaca memahami apa yang disampaikan pada Bab IV dan V."

"Background: The prevalence of Helicobacter pylori (H. pylori) infection in the world is quite high, especially in developing countries. Usually the patient shows no specific symptoms and chronic gastritis therefore becomes chronically infected The complication of the injéction is the development of peptic ulcer; which is a predisposing factor for gastric carcinoma. Early diagnosis is an important step to avoid these complications by providing immediate accurate therapy. Methods: In this study the CLO, MIU (Motility Indole Urease) tests and culture were conducted on 131 biopsy samples of the stomach antrum mucous tissue taken from chronic dyspepsia patients from several hospitals in Jakarta. In the CLO test, biopsy tissue was put in a small well agar to be incubated at room temperature. In the MIU test the biopsy tissue sample was submerged in the small MlU tube agar with a depth of approximately 2/3 rds from the surface, and then incubated at room temperature. Another piece of biopsy tissue was cultured micro-aerophylicalty The CLO and MlU tests are considered positive if the color changes from yellow to red and are considered negative if there is no color change within 24 hours. Results: Compared to culture, the CLO test demonstrated 38% sensitivity; 96% specificity, 94% positive predictive value and 52% negative predictive value, whereas the results of the MIU test against culture method showed 76% sensitivity 89% specificity 88% positive predictive value, and 78% negative predictive value. Conclusion: The MIU test that showed high sensitivity and specyficity and thus could be further developed as an alternative diagnostic method for H. pylori infection."

"Latar belakang: Sepsis sulit dibedakan dengan respon inflamasi non infeksi secara klinis dan kultur darah yang merupakan gold standar diagnosis sepsis memiliki banyak keterbatasan. Presepsin merupakan suatu biomarker baru namun belum banyak data tentang efektifitas penggunaanya pada anak.Tujuan: Mengetahui nilai diagnostik dan prognostik presepsin dibandingkan dengan leukosit, PCT dan CRP pada pasien anak yang dicurigai sepsis

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Method: The latitude cut study was conducted during March-December 2020 at RSCM Jakarta on 56 patients aged 2 months - 10 years with suspicion of sepsis Diagnosis of sepsis is established based on the criteria of sepsis-3 and blood culture. Biomarker examination and PELOD-2 score are performed at the beginning and after 72 hours, mortality assessment is conducted on day 7. Presepsin levels are checked using the PATHFAST® method.

Result: The median value of precessine levels in the proven sepsis group (1183 pg/ml was higher than that of the unproven group of sepsis (369 pg/ml, p=0.001). Precessine has a good diagnostic value (AUC of 0.862), with a cut of 711 pg/ml having a sensitivity of 75.8%, specificity of 82.6%, positive guess value of 86.2% and negative guess value of 70.4%, better than leukocytes, PCT, and CRP. Presepsin levels increased linearly with the severity of sepsis and were moderately correlated with PELOD-2 scores (r=0.548; p=0.001). Survival analysis showed precessine levels of ≥ 1,250 pg/ml were significantly associated with early mortality (HR 6.31; 95%CI; 1.67-23.83; p=0.007). Presepsin levels after 72 hours of antibiotic therapy decreased significantly in the improved sepsis group and increased in the worsening sepsis group.

Inference: Presepsin is a reliable biomarker and can be used to help diagnose sepsis, predict severity, death and evaluate therapies in tertiary hospital services."

"Latar Belakang: Uveitis infeksi di Indonesia berkisar antara 30-60% dari total kasus uveitis. Identifikasi patogen etiologi infeksi sangat penting agar dapat diberikan terapi antimikroba yang sesuai dengan segera sehingga komplikasi kebutaan dapat diminimalisir. Dewasa ini, perkembangan teknologi biologi molekuler menggunakan metode Polymerase Chain Reaction (PCR) untuk deteksi uveitis infeksi sedang berkembang pesat.

Tujuan: Melakukan uji validasi diagnostik pada metode PCR Multiplex dibandingkan dengan metode PCR Tunggal.

Metodologi: Uji diagnostik untuk menentukan sensitivitas dan spesifisitas dari alat Real-Time PCR Multiplex terhadap PCR Tunggal dari spesimen cairan intraokular humor akuos yang diambil dari parasentesis bilik mata depan. Dilakukan pemeriksaan PCR terhadap patogen Mycobacterium tuberculosis (M.tuberculosis), Toxoplasma gondii (T.gondii), Herpes Simplex Virus (HSV), Varicella Zoster Virus (VZV), Cytomegalovirus, dan Treponema pallidum.

Hasil: Dilakukan analisis uji diagnostik pada 46 subjek penelitian. Didapatkan hasil sensitivitas sebesar 57.14% dan spesifisitas sebesar 100%. Positivity rate terbanyak didapatkan untuk patogen VZV (n=4), dan tidak didapatkan hasil positif terhadap deteksi patogen M.tuberculosis. Patogen T.pallidum berhasil dideteksi sebanyak 4.34% (n=2) oleh PCR Multiplex.

Kesimpulan: Metode PCR Multiplex pada penelitian ini memiliki sensitivitas yang rendah dengan spesifisitas yang tinggi. Hasil positif pada PCR Multiplex dapat bermanfaat untuk mendiagnosis pasien dengan uveitis infeksi.

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Background: In Indonesia, infectious uveitis represents 30-60% of the country’s total uveitis cases. The identification of etiological pathogens is imperative to immediately select and administer the appropriate antimicrobial therapy in infectious uveitis, thereby complications of blindness can be minimized. Currently, the development of molecular biology technology using the Polymerase Chain Reaction (PCR) method for detection of infectious uveitis pathogens is growing rapidly.

Objective: To compare the diagnostic validation test results of the Multiplex PCR method and Single PCR method.

Method: Diagnostic test to determine the sensitivity and specificity of the Multiplex Real-Time PCR device against the Single PCR of aqueous humor intraocular fluid specimens taken from anterior chamber paracentesis. PCR examinations were carried out to identify the pathogens of Mycobacterium tuberculosis (M.tuberculosis), Toxoplasma gondii (T.gondii), Herpes Simplex Virus (HSV), Varicella Zoster Virus (VZV), Cytomegalovirus, dan Treponema pallidum.

Result: A diagnostic test analysis was performed on 46 study subjects. The results obtained 57.14% sensitivity and 100% specificity. Highest positivity rate was obtained for VZV pathogens, while positive results were not obtained for M.tuberculosis. There were 4.34% of subjects (n = 2) of T. pallidum were detected by PCR Multiplex. 

Conclusion: The PCR Multiplex method in this study has low sensitivity with high specificity. A positive result on Multiplex PCR can be useful for diagnosing patients with infectious uveitis."