Anis Karuniawati
Departemen Mikrobiologi Klinik Fakultas Kedokteran, Universitas Indonesia

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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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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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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.
Construction of pcDNA3.1 Vector Encoding RpfD Gene of Mycobacterium tuberculosis Rakhmawati, Aprilia; Rukmana, Andriansjah; Karuniawati, Anis
Makara Journal of Science
Publisher : UI Scholars Hub

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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.
Efficacy of Tuberculosis Vaccine Candidate pcDNA3.1-rpfB in Inhibiting the Growth of Mycobacterium tuberculosis In Vitro with Mycobacterial Growth Inhibition Assay Pujilestari, Ratih; Rukmana, Andriansjah; Karuniawati, Anis
Makara Journal of Science Vol. 26, No. 1
Publisher : UI Scholars Hub

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Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb). Bacille Calmette-Guérin (BCG) is the only licensed vaccine against TB, and it is effective in children but not in adults. The Vaccine Research Team, Department of Microbiology FKUI has developed a DNA-based TB vaccine candidate pcDNA3.1-rpfB. This candidate induces immune responses in mice, but its potency is unknown. The gold standard for potency testing of TB vaccine is the challenge method. The BSL3 animal laboratory for the challenge method is currently unavailable at FKUI. Therefore, mycobacterial growth inhibition assay (MGIA) was used as a preliminary test before the in vivo challenge test was conducted. The principle of MGIA is to reculture Mtb in a Mycobacteria Growth Indicator Tube (MGITTM) from co-cultured Mtb with mammalian cells that have been previously treated with pcDNA3.1-rpfB, pcDNA3.1 (negative control), and BCG (positive control). MGITTM shows the time to positivity, which is the time that has lapsed until a positive growth of Mtb is detected. In addition, measurements of interferon (IFN)γ levels by enzyme-linked immunosorbent assay were carried out. This study concluded that pcDNA3.1-rpfB can inhibit the growth of Mtb in vitro and showed no statistical difference from BCG. The IFNγ levels from co-culturing did not correlate with the level of inhibition of the growth of Mtb in vitro.
The Role of Clinical Sign and The Added Value of Procalcitonin in Determining The Existance of Infection in The Treated Diabetic Foot Ulcer Aswar, Andra; Yunir, Em; Karuniawati, Anis; Harimurti, Kuntjoro
Jurnal Penyakit Dalam Indonesia Vol. 5, No. 2
Publisher : UI Scholars Hub

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Correlation between Proper Antibiotics Administration to Community Acquired Pneumonia Patients Based on Alur Gyssens with Clinical Response Rumende, Cleopas Martin; Chen, Lie Khie; Karuniawati, Anis; Bratanata, Joyce; Falasiva, Rezyta; Sitorus, Truly Panca; Susanto, Erwin Christian
Jurnal Penyakit Dalam Indonesia Vol. 6, No. 2
Publisher : UI Scholars Hub

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Introduction. The proper of antibiotics treatment will increase the improvement of the patient with community-acquired pneumonia (CAP) and also counteract antimicrobial resistance. Community-Acquired Pneumonia is commonly found in Indonesia with an incidence of 2.4%. This study aimed to determine the proper of antibiotic selection for CAP based on IDSA-ATS 2007 guidelines with Gyssens method and evaluate the relationship between the proper of antibiotic treatment with the clinical response of patients. Methods. A retrospective cohort study was conducted with secondary data from the previous study. Study was conducted in three hospitals in Jakarta, namely Cipto Mangunkusumo General Hospital, Budhi Asih Hospital, and Gatot Subroto Hospital between September 2016 and November 2017. Results. From 151 patients, the appropriate of the first antibiotic treatment, including category 0 and 1 was found in 38 patients (25.2%), while for inappropriate one (category II-V) was in 113 patients (74.8%) mainly due too short antibiotic treatment. For the second antibiotic given to 54 patients, appropriate treatment was found in 15 patients (9.9%). There was no correlation between the accuracy of both antibiotics treatment in CAP based on Gyssens method with clinical response of the patients (first antibiotic treatment: RR=0,970 (95% CI: 0,80 – 1,16), p=0,738; second antibiotic treatment: RR=1,194 (95% CI: 0,648 – 2,20), p=1,00). Conclusion. There was no correlation between the accuracy of antibiotics treatment in CAP based on Gyssens method with clinical response of the patients.
Performance of Xpert MTB/RIF by Using Gastric Lavage Samplein The Diagnosis of Pulmonary Tuberculosis in HIV Patients withSuspected Pulmonary Tuberculosis Mboeik, Manuel Lamberto Willem; Pitoyo, Ceva Wicaksono; Karjadi, Teguh Harjono; Karuniawati, Anis; Dewiasty, Esthika
Jurnal Penyakit Dalam Indonesia Vol. 5, No. 1
Publisher : UI Scholars Hub

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Introduction. Xpert MTB/RIF test on gastric lavage sample may improve pulmonary tuberculosis (TB) case finding, but its performance as a diagnostic test for pulmonary TB in HIV-infected adults patients has not been studied widely, including in Indonesia. This study aims to determine the performance of the Xpert MTB/RIF test in diagnosing pulmonary tuberculosis 30 | Jurnal Penyakit Dalam Indonesia | Vol. 5, No. 1 | Maret 2018 Manuel Lamberto Willem Mboeik, Ceva Wicaksono Pitoyo, Teguh Harjono Karjadi, Anis Karuniawati, Esthika Dewiasty among HIV-infected adult patients with suspected pulmonary tuberculosis by using gastric lavage sample. Method. A cross-sectional study was conducted on adult HIV-infected patients with suspected pulmonary tuberculosis in Cipto Mangunkusumo hospital Jakarta between February 2016 and June 2017. Gastric lavage sample and sputum sample collection were performed for Xpert MTB/RIF test and acif fast bacilli (AFB) smear examination. Results. Among total of 117 subjects, 81 (69,2%) subjects were unable to expectorate sputum. M tuberculosis was detected on Xpert MTB/RIF using gastric lavage samples in 34 (29,1%) subjects, AFB in 4 of 36 sputum samples and 2 of 117 gastric lavage samples. The drug sensitivity test on all 34 Xpert MTB/RIF positive subjects showed no rifampicin resistance. Fifty-one (43.6%) subjects were diagnosed as clinical pulmonary tuberculosis, 26 subjects among them were positive on Xpert MTB/ RIF assay, 4 subjects were smear sputum positive and 1 subject was gastric lavage smear positive. Using clinical diagnosis of pulmonary TB as gold standard, a single Xpert MTB/RIF assay using gastric lavage sample showed sensitivity of 50.98% (95% CI: 36.6-65.25), specificity 87.88% (95% CI: 77,1-94,62), positive predictive value 76,47% (95%CI: 61,67-86,78) and negative predictive value 69,88% (95% CI: 63,36-75.68). Conclusion. Xpert MTB/RIF performed on gastric lavage samples showed sensitivity of 50.98% and specificity 87.88% in clinical diagnosis of pulmonary tuberculosis in HIV-positive patients with pulmonary tuberculosis.