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Optimizing real-time PCR method to detect Leptospira spp. in human blood and urine specimens Karuniawati, Anis; Yasmon, Andi; Ningsih, Ika
Medical Journal of Indonesia Vol 21, No 1 (2012): February
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (925.644 KB) | DOI: 10.13181/mji.v21i1.472

Abstract

Background: Leptospirosis is an acute infectious disease in humans caused by Leptospira spp. and classified as a zoonosis. Clinical symptoms of leptospirosis are nonspecific and the current available laboratory method for detecting Leptospira spp. is difficult, which resulted to the misdiagnosis of this disease. Therefore, the rapid and accurate method is needed to diagnose the disease. This study was aimed to optimize molecular diagnostic test using real-time PCR assay as a rapid, sensitive and specific method for the detection of pathogenic Leptospira spp. in humans.Methods: Bacterial DNA was extracted by DNA extraction kit according to the manufacturer’s instructions. Primers and probes used in this study was based on previous and published research. The assay is performed using PCR-IQTM5, iCycler Multicolor real-time PCR detection system. Specificity of the primer used was evaluated towards some bacterial pathogens.Results: Limit detection of the DNA was 0.375 fg/ml and the primers used does not cross-react with the genomes of the pathogens tested. Limit detection of DNA in blood is 150 fg/μl, and in urine is 1470 fg/μl.Conclusion: Real-time PCR test is a rapid and accurate method for detecting pathogenic Leptospira spp. in human specimens. Further research is needed to determine the sensitivity and specificity of real-time PCR tests compared with other diagnostic methods in clinical settings. (Med J Indones 2012;21:13-7)Keywords: Leptospirosis, Leptospira, optimization, real-time PCR
PENGARUH FINGER PAINTING TERHADAP KEMAMPUAN MENGENAL KONSEP WARNA PADA ANAK KELOMPOK A Karuniawati, Anis
PAUD Teratai Vol 6, No 3 (2017): Vol 6 No 3 Edisi Yudisum Sep 2017
Publisher : PAUD Teratai

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Abstract

Abstrak   Penelitian Pre-Experimental Design ini bertujuan untuk mengkaji ada atau tidaknya pengaruh finger painting terhadap kemampuan mengenal konsep warna pada anak kelompok A di TK Dharma Wanita Krikilan III Driyorejo Gresik. Populasi penelitian ini adalah anak kelompok A di TK Dharma Wanita Krikilan III Driyorejo Gresik dengan sampel 13 anak kelompok A. Teknik pengumpulan data menggunakan observasi dan dokumentasi. Teknik analisis data penelitian ini menggunakan Wilcoxon Match Pairs Test dengan rumus Thitung < Ttabel, dengan menggunakan taraf signifikan 5%. Jika Thitung lebih kecil daripada Ttabel, maka Ho ditolak dan Ha diterima. Berdasarkan hasil pengolahan data diperoleh Thitung = 0 dan Ttabel untuk N= 13 dengan taraf signifikan 5%  diperoleh sebesar 17, maka 0 < 17. Dengan demikian, maka Ho ditolak dan Ha diterima.  Jadi, dapat disimpulkan bahwa finger painting berpengaruh terhadap kemampuan mengenal konsep warna pada anak kelompok A di TK Dharma Wanita Krikilan III Driyorejo Gresik. Kata kunci: finger painting, konsep warna.   Abstract Research Pre Experimental Design aims to examine influence of finger painting to the ability of recognizing the concept of color in childhood in group A TK Dharma Wanita Krikilan III Driyorejo Gresik. The population is children in group A TK Dharma Wanita  Krikilan III Driyorejo Gresik with  a sample of 13 children. Techniques data collection using observation and documentation. Technique of data analysis of this research use wilcoxon match pairs test with formula Tcount < Ttable, with significant level 5%. The result of data analysis shows that Tcount = 0, while Ttable  with N=13 obtained of 17, then 0 < 17. If the price Tcount smaller than Ttable , Ho is rejected and Ha accepted. So,it can be concluded that finger painting is influential on the ability to recognizing of concept color in group A TK Dharma Wanita  Krikilan III Driyorejo Gresik. Keywords: finger painting, color concept
Epidemiology of Microorganisms in intraabdominal infection/complicated intraabdominal infections in six centers of surgical care in Indonesia: A preliminary study Moenadjat, Yefta; Lalisang, Toar JM.; Saunar, Rofy S.; Usman, Nurhayat; Handaya, Adeodatus Y.; Iswanto, J.; Nasution, Safruddin; Karuniawati, Anis; Loho, Tony; Widyahening, Indah S.
The New Ropanasuri Journal of Surgery
Publisher : UI Scholars Hub

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Abstract

Introduction. Data of complicated intraabdominal infections (cIAI) and the epidemiology of causative microorganisms which is Indonesian characteristics is required to develop a guideline. Thus, a preliminary study run to find out such characteristics. Method. Data of subjects with cIAI managed in six centers of teaching hospital in Indonesia in period of 2015–2016 were collected. Those data of source of infection, the epidemiology of microorganism and susceptibility of antibiotics were descriptively provided. Results. Source of infection were perforated appendicitis (26.64%), perforated gastric and duodenal ulcer (22.70%), small bowel perforation (11.84%), large bowel perforation (13.16%), postoperative (9.54%), and others (16.2%). Escherichia coli and Klebsiella pneumonia were the most microorganisms found in the pus specimen. The sensitivity of Escherichia coli and Klebsiella pneumonia to cephalosporins were in range of 14.1– 42% and 28.7–35.6%, respectively. Conclusion. Perforated appendicitis, perforated gastric and duodenal ulcer, small bowel perforation, large bowel perforation, and postoperative in sequent are the main causal of cIAIin Indonesia. The epidemiology predominated by Gram negative, particularly Escherichia coli and Klebsiella pneumonia.
Identifikasi Brucella abortus Isolat Lokal dengan Brucella abortus Strain Specific-Polymerase Chain Reaction (IDENTIFICATION OF LOCAL ISOLATES OF BRUCELLA ABORTUS USING BRUCELLA ABORTUS STRAIN SPECIFIC-POLYMERASE CHAIN REACTION ASSAY) Susan Maphilindawati Noor; Pratiwi Pujilestari Sudarmono; Asmarani Kusumawati; Anis Karuniawati
Jurnal Veteriner Vol 15 No 3 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Brucella abortus Strain Specific-Polymerase Chain Reaction (BaSS-PCR) is a single multiplex PCRtechnique which able to identify and differentiate between Brucella abortus field strains (biovar 1, 2, and4), B. abortus vaccine strains, Brucella species, and non-Brucella species. In this study, BaSS-PCR wasapplied to identify local isolates of B. abortus in order to investigate the B. abortus strains that infectedcattle in Indonesia. Fifty local strains of B.abortus isolated from infected cattle in Java (Jakarta andBandung), South Sulawesi (Maros), East Nusa Tenggara (Kupang and Belu) were used in this study. TheDNA bands were observed by agarose gel in the presence of ethidium bromide. Identification was performedbased on the size and number of DNA products amplified by PCR from each isolates. The results showedthat the 50 isolates were of B. abortus field strains. This finding showed that the cause of bovine brucellosisin Indonesia is B. abortus field strains.
Epithelial Cells Count and the Ratio of Leukocytes and Epithelial Cells as the Criteria to Determine Qualified Specimen for Community-Acquired Pneumonia (CAP)-causing Pathogens Identification Ade Dharmawan; Anis Karuniawati; Pratiwi Pudjilestari Sudarmono; Delly Chipta Lestari; Cleopas Martin Rumende
The Indonesian Biomedical Journal Vol 12, No 1 (2020)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v12i1.873

Abstract

BACKGROUND: Community-acquired pneumonia (CAP) is the most common infectious with serious rate of morbidity and mortality. Recent conventional method only described 30-50% of CAP etiology. Sputum specimen quality assessment is important to obtain an accessible CAP-causing pathogens identification.METHODS: This was a prospective descriptive study involving 100 specimens from CAP-diagnosed subjects in Budhi Asih Regional General Hospital inpatien tcare. We assessed three gram-staining criteria for specimen quality determination, and continued by bacterial identification.RESULTS: All specimens were qualified according to criteria II, while only 94 and 96 specimens were qualified according to criteria I and III, respectively. Sixty-five specimens could be identified by culture and pneumoCLART polymerase chain reaction (PCR) examination, and the 35 specimens remained unknown. Ten out of those 35 specimens were positive after analyzed by Acid-fast Bacilli (AFB) test. The pathogens we identified including Klebsiella pneumoniae (29.6%), Acinetobacter baumanii (10.2%), Enterobacter cloacae (4.6%), Pseudomonas aeruginosa (4.6%), Staphyloccocus aureus (4.6%), Moraxella catarrhalis (3.7%), Enterobacter aerogenes (2.8%), Escherichia coli (2.8%), Streptococcus pneumoniae (1.9%), Mycoplasma pneumoniae (1.9%) and Citrobacter koseri (0.9%).CONCLUSION: There were no significant differences among the three criteria for sputum specimen quality assessment, based on culture and pneumoCLART examination. We suggest that criteria II could be used to avoid many specimen rejections while good quality specimens still attained for accessible bacteria identification.KEYWORDS: community-acquired pneumonia, sputum, gram stain, pathogens, bacteria
Gene Families of AmpC-producing Enterobacteriaceae Present in the Intensive Care Unit of Cipto Mangunkusumo Hospital Jakarta Lucky Hartati Moehario; Thomas Robertus; Anis Karuniawati; Rudyanto Sedono; Delly Chipta Lestari; Andi Yasmon
The Indonesian Biomedical Journal Vol 11, No 1 (2019)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v11i1.552

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BACKGROUND: Antibiotic resistance has become a worldwide problem. Among Asia countries, Indonesia has high prevalence of multi-drug resistant organisms mainly due to Gram-negative bacilli Enterobacteriaceae. This study aimed to find out whether gene family of AmpC and AmpC/ESBL-producing Enterobacteriaceae were present in the Intensive Care Unit (ICU) of Cipto Mangunkusumo Hospital, Jakarta, Indonesia.METHODS: Specimens were obtained from several body sites of adult patients with infection hospitalised in ICU of Cipto Mangunkusumo Hospital. VITEK®2 was used to identify the microorganisms. Antibiotic susceptibility tests were conducted using VITEK®2 and disc diffusion technique according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Double disc synergy (DDS) test method was employed to detect AmpC activity. Gene families of ampC were identified using multiplex polymerase chain reaction (PCR).RESULTS: Forty five isolates were identified as putative AmpC, extended-Spectrum β-lactamases (ESBL) and AmpC/ESBL-producing Enterobacteriaceae. Klebsiella pneumoniae (n=32) were predominant, followed by Escherichia coli (n=6), Enterobacter cloacae (n=5) and Enterobacter aerogenes (n=2). AmpC activity was detected in 9 isolates, in which 4 isolates were AmpC producing and 5 isolates were AmpC/ESBL. In vitro, AmpC-producing Enterobacteriaceae showed good susceptibility to many antibiotic tested, while those of AmpC/ESBL-producing only to Amikacin. The gene families of ampC were DHA, EBC and CIT identified from 6 isolates.CONCLUSION: DHA, EBC and CIT gene families were identified from AmpC and AmpC/ESBL-producing Enterobacteriaceae in the ICU of Cipto Mangunkusumo Hospital. While the AmpC-producing was still susceptible to almost all antibiotics tested, the AmpC/ESBL-producing showed resistant except for Amikacin.KEYWORDS: Enterobacteriaceae, β-lactamases, AmpC, ESBL
Comparison of minimal inhibitory and bactericidal capacity of oral penicillin V with benzathine penicillin G to Streptococcus beta--hemolyticus group A in children with rheumatic heart disease Burhanuddin Iskandar; Bambang Madiyono; Sudigdo Sastroasmoro; Sukman T. Putra; Mulyadi M. Djer; Anis Karuniawati
Paediatrica Indonesiana Vol 48 No 3 (2008): May 2008
Publisher : Indonesian Pediatric Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (233.281 KB) | DOI: 10.14238/pi48.3.2008.152-5

Abstract

Background Injection ofbenzatine penicillin G (BPG) every 28days is still the drug of choice for secondary prevention of rheu-matic heart disease (RHD). BPG sometimes poses problems dueto pain at the injection site, possible anaphylaxis, and is not alwaysavailable. Some centers choose oral penicillin over BPG.Objectives To compare minimal inhibitory capacity (MIC) andminimal bactericidal capacity (MBC) of oral penicillin V serumwith those of BPG among SGA infected RHD.Methods This was a clinical trial with crossover design study tocompare MIC of penicillin V and BPG. Outcome measures wereMIC and MBC. Statistical analysis was performed using pairedt-test and wilcoxon test.Result There were 32 subjects consisted of 17 males and 15females. The mean value of MIC and MBC serum of penicillinV were 0.031 and 0.125. The mean value of MIC and MBCserum of BPG3 were 0.094 and 0.031. Respectively the MICof penicillin V was similar to that of BPGy The mean value ofMIC and MBC of BPG4 were 0.125 and 0.250. Respectively theMIC of penicillin V was significantly higher than that of BPG 4.The MBC of penicillin V was significantly higher than that ofBPG 4. The MIC ofBPG 3 was similar to that ofBPG 4• The MBCof BPG 3 was similar to that of BPG 4.Conclusions The MIC of penicillin V was similar to that ofBPG 3,the MBC of oral penicillin V was higher than that ofBPG 3• TheMIC and MBC of penicillin V was higher than those of BPG 4.
PREVALENSI ISOLAT MRSA PENGHASIL PANTON-VALENTINE LEUKOCIDIN PADA PASIEN ICU RUMAH SAKIT TERSIER Linosefa Linosefa; Delly Chipta Lestari; Ardiana Kusumaningrum; Anis Karuniawati; Andi Yasmon
Majalah Kedokteran Andalas Vol 39, No 1 (2016): Published in April 2016
Publisher : Faculty of Medicine, Universitas Andalas

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (529.517 KB) | DOI: 10.22338/mka.v39.i1.p1-10.2016

Abstract

Penilitian ini bertujuan untuk mengetahui prevalensi MRSA penghasil Panton-Valentine leukocidin (PVL) dan pola kepekaannya. Sampel penelitian adalah isolat MRSA dari 315 pasien Rumah Sakit Umum Pusat Nasional Cipto Mangunkusumo (RSUPNCM) selama tahun 2011 dan 2014, dengan melakukan identifikasi, uji kepekaan dan uji molekuler terhadap isolat tersebut. Penelitian ini menunjukkan sebanyak 59% dari koloni MRSA yang ditemukan masih sensitif terhadap antibiotik golongan selain β-laktam, sehingga masih dapat diduga sebagai community-associated MRSA (CA-MRSA). CA-MRSA sepertinya mulai ditransmisikan di fasilitas kesehatan. Uji molekuler terhadap isolat MRSA memberikan hasil 8,3% isolat MRSA menghasilkan PVL. Berdasarkan tipe pola kepekaannya isolat MRSA penghasil PVL tersebut masih dapat digolongkan sebagai CA-MRSA. MRSA penghasil PVL ditemukan di RSUPNCM sebagai kolonisasi. Surveilan perlu dilakukan untuk memahami interaksi antara MRSA di komunitas dan rumah sakit, terutama untuk mengurangi transmisi di fasilitas kesehatan.
Necrotizing Fasciitis in a Leprosy Patient Nie Nie; Augustine Natasha; Sweety Pribadi; Yulia R. Saharman; Chairunissa T. Rizal; Anis Karuniawati; Pratiwi Sudarmono
eJournal Kedokteran Indonesia Vol 10, No. 1 - April 2022
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (570.017 KB) | DOI: 10.23886/ejki.10.51.77-80

Abstract

Leprosy is an infection caused by Mycobacterium leprae. Disability after the infection is common  and necrotizing fasciitis could deteriorate the patient current condition. We present the case of necrotizing fasciitis of the right limb inpatient with tarsal disintegration and plantar ulcer due to previous leprosy infection.  The microbiology culture was inadequate in the early stage of management, which delayed the definitive  antibiotic for patient and the progressive necrotic infections were uncontrolled. Amputation was done to save the patient’s life and the latter microbiology culture was able to determine the definitive antibiotic therapy. This report highlights the needs of disability management and infection control for leprosy patient after the treatment is completed.
Construction of pcDNA3.1 Vector Encoding RpfD Gene of Mycobacterium tuberculosis Rakhmawati, Aprilia; Rukmana, Andriansjah; Karuniawati, Anis
Makara Journal of Science Vol. 22, No. 3
Publisher : UI Scholars Hub

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Abstract

Tuberculosis (TB) is an infectious diseasecaused by Mycobacterium tuberculosis (M. tuberculosis). TB is still a major health problem. The Bacillus Calmette-Guérin (BCG) vaccineis the only one available for TB and is known to confer variable levels of protection. Because of thisvariability, a new vaccine is needed to control TB. Proteins secreted by M.tuberculosisare known to induce protective immunity. Within the genome of M. tuberculosis, there is a family of proteins called resuscitation promoting factor (Rpf), which playsa role in the reactivation of M. tuberculosis. RpfD is amember of the Rpf family that has been shown to be immunogenic, makingitsuitable for use as a TB vaccine. The rpfD gene of the M. tuberculosis Beijing strain from the bacterial stock of the Department of Microbiologyat the Medical Facultyof theUniversitas Indonesia was amplified using polymerase chain reaction (PCR) and then insertedintothemammalian expression vector pcDNA3.1(+). Then, the pcDNA3.1(+)-rpfD vector was transformed to Escherichia coli DH5α. A 465-bp target fragment was obtained, and the accuracy ofthecloning was confirmed using colony PCR, restriction enzyme digestion, and sequencing. We expect that this recombinant plasmid will induce immunity in future animal models and thus will prove itself to be a candidate for an M. tuberculosis vaccine.
Co-Authors . Andriansjah Ade Dharmawan Amalia Almira Amarila Malik Amin Soebandrio Andi Annisa Rusyda Khafiyani Andi Yasmon Andra Aswar Andra Aswar, Andra Angky Budianti Anwar S. Ibrahim Aprilia Rakhmawati Ardiana Kusumaningrum Ari Fahrial Syam Ari Prayitno Aria Kekalih Aryo Tedjo Asmarani Kusumawati Augustine Natasha Aulia Rizki Bachti Alisyahbana Badriul Hegar Badriul Hegar Bambang Madiyono Burhanuddin Iskandar Ceva W. Pitoyo Chairunissa T. Rizal Cleopas Martin Rumende Cleopas Martin Rumende Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Dimas Seto Prasetyo Dwiana Ocviyanti E. Risdiyani Em Yunir Em Yunir, Em Erwin Christian Susanto Esthika Dewiasty Esthika Dewiasty, Esthika Falasiva, Rezyta Fariz Nurwidya Febrianti, Tati Febriyana, Dwi Futihati Ruhama Zulfa Gestina Aliska Gortap Sihotang Hana Paraswati Putri Handaya, Adeodatus Y. Hindra Irawan Satari Hindra Irawan Satari Ida Parwati Ika Ningsih Indah S. Widyahening Iswanto, J. Joyce Bratanata Joyce Bratanata, Joyce Kambang Sariadji Khie Chen Kuntjoro Harimurti Lalisang, Toar JM. Linosefa Linosefa Manuel Lamberto Willem Mboeik Martin Hartiningsih Mboeik, Manuel Lamberto Willem Moehario, Lucky Hartati Muhammad K Azwar, Muhammad K Mulya Rahma Karyanti, Mulya Rahma Mulyadi M. Djer Muziasari, Windi Nabila Maudy Salma Nasution, Safruddin Nelly Puspandari Nie Nie Nina Dwi Putri Nuraini I Susanti Permatasari, Tri Wijayanti Pratama Wicaksana Pratiwi Pujilestari Sudarmono Pratiwi Sudharmono Prawoto Prawoto Prawoto Prawoto Pujilestari, Ratih Purwantyastuti Purwantyastuti Puspandasari, Nelly Rakhmawati, Aprilia Resti H. Lestari Reynaldo Reynaldo Reynaldo Reynaldo, Reynaldo Rezyta Falasiva Riamin Sitorus Rianto Setiabudi, Rianto Rida Tiffarent Rosdiana Irawati Rudyanto Sedono Rudyanto Sedono Rudyanto Sedono, Rudyanto S. Nilawati Santya Fatma Dewi Sari Wiraswasty Saunar, Rofy S. Shindy Claudya Aprianti Simanjuntak, Glory Gelarich Sitorus, Truly Panca SJATHA, FITHRIYAH Subari A. Riyanto Sudigdo Sastroasmoro Sukman T. Putra Sunarno Sunarno Susan M. Noor Susan Maphilindawati Noor Susanti, Nuraini I Susanti, Nuraini I Susanto, Erwin Christian Sweety Pribadi SYADZA RHIZKY PUTRI AKHMAD T. M. Sudiro Teguh Harjono Karjadi Teguh Harjono Karjadi, Teguh Harjono TETSUHIRO MATSUZAWA Thomas Robertus Tjampakasari, Conny Riana TOHRU GONOI Tony Loho Trisni U Dewi, Trisni U Truely Panca Sitorus Usman, Nurhayat Weaver, Tom Wia Melia Windi Muziasari Yefta Moenadjat Yeva Rosana Yulia R. Saharman Yulia Rosa Saharman