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Multiplex nested polymerase chain reaction for Treponema pallidum using blood is more sensitive than using serum Effendi, Ida; Rosana, Yeva; Yasmon, Andi; Indriatmi, Wresti
Universa Medicina Vol 37, No 1 (2018)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2018.v37.75-84

Abstract

BackgroundSyphilis is a multistage disease transmitted primarily through sexual intercourse. Nowadays, the polymerase chain reaction (PCR) test for Treponema pallidum has been widely used and is expected to overcome problems in diagnostic tests for syphilis. The Treponema pallidum PCR is influenced by type of specimens, PCR methods and target genes. This study aimed to assess the use of blood and serum in multiplex nested PCR for Treponema pallidum, targeting the 23S rRNA.MethodsA cross-sectional study was conducted from April 2015 - April 2016. Sampling was carried out consecutively among patients with clinical features of secondary syphilis who came to Sexually Transmitted Disease (STD) clinics in Jakarta. All sera were also tested with Rapid Plasma Reagin (RPR) and Treponema pallidum Hemagglutination Assay (TPHA) assay, which was considered as the gold standard for this study. We determined the sensitivity and specificity of the multiplex nested PCR for Treponema pallidum using blood and serum.ResultsPCR test was performed on 122 clinical specimens (61 blood and 61 serum). The positive results of PCR test on blood was 22.95% and serum was 6.56%, while the positive results of serology was 68.85%. The sensitivity of Treponema pallidum multiplex nested PCR on blood was 30.95% compared to serum 9.52% (p=0.006). PCR test on blood is able to detect 3.25 times higher than serum. ConclusionThe use of blood has a higher proportion of positives compared to serum in Treponema pallidum multiplex nested PCR using 23S rRNA target gene.
Microscopic examination using negative staining for rapid diagnosis of syphilis Rosana, Yeva; Effendi, Ida; Indriatmi, Wresti; Yasmon, Andi
Universa Medicina Vol. 41 No. 1 (2022)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2022.v41.64-70

Abstract

BACKGROUNDSyphilis is a global health problem, especially in developing countries including Indonesia. Treponema pallidum, the etiologic agent of syphilis, cannot be cultured in vitro. Syphilis has several clinical manifestations, making laboratory testing a very important aspect of diagnosis. Microscopic examination may support the diagnosis but is rarely used in Indonesia. The aim of this study was to evaluate negative staining using the light microscope to detect T. pallidum in syphilitic lesions. METHODSA cross-sectional study was conducted involving 27 subjects who came to several dermato-venereology clinics in Jakarta. Exudates were collected from genital ulcers, condylomata lata, and dry mucocutaneous rash on palms and soles of syphilis patients. Negative staining using one drop of Indian ink was used to examine for treponemas under the light microscope at 10x100 magnification. RESULTSMicroscopic examination using negative staining showed a few clusters of small and spiral shaped bacteria. Of the 39 specimens from 27 subjects, microscopic examinations were successfully done on 10 specimens. Observations could only be conducted on 5 specimens, 3 (60.0%) of which showed the morphology of spirochetes. This examination is the easiest method for detecting the bacteria. Moreover, the bacteria that were isolated from painless genital ulcers could be observed more clearly than those from erythematous maculopapular lesions. CONCLUSIONTreponema pallidum was successfully detected by microscopic examination in all moist lesions, but was difficult to detect in dry lesions. Negative staining under the light microscope appears to be simple, affordable, and available in most microbiology laboratories in Indonesia.
Development of a SYBR Green real-time PCR-based assay system for detection of Neisseria gonorrhoeae Andi Yasmon; Rela Febriani; Louisa Ivana Utami; Fithriyah Fithriyah; Yeva Rosana; Fera Ibrahim; Pratiwi Sudarmono
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 54, No 1 (2022)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19106/JMedSci005401202201

Abstract

Diagnosis of Neisseria gonorrhoeae infection is needed for patient therapy and for reducing this bacterial transmission in the population. The culture method is a gold standard method for N. gonorrhoeae detection, however it has low sensitivity. Among molecular methods with high sensitivity and specificity, SYBR Green real-time PCR is the potential method for N. gonorrhoeae detection. In this study, we developed an SYBR Green real-time PCR-based system assay for N. gonorrhoeae detection. Several PCR conditions were optimized and analyzed including primer annealing temperature, DNA template volume, the limit of detection (LoD), cross-reaction with others (bacteria, viruses, fungus, protozoa), and quality assurance. The results showed that the annealing temperature and DNA template volume were 60oC and 5 µL, respectively. The LoD was 29 DNA copies corresponding to 3 bacterial cells per reaction. No cross-reaction was detected for other bacteria, viruses, fungus and protozoa. The external quality assurances enrolled in 2019 and 2021 showed 100% concordance. The preliminary testing for clinical samples was also 100% concordance. In conclusion, the SYBR Green real-time PCR-based system assay developed in this study is promising for application in clinical laboratories.
The antifungal susceptibility of Candida albicans isolated from HIV/AIDS patients Sri Rezeki; Siti Aliyah Pradono; Gus Permana Subita; Yeva Rosana; S. Sunnati; Basri A. Gani
Dental Journal (Majalah Kedokteran Gigi) Vol. 54 No. 2 (2021): June 2021
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/j.djmkg.v54.i2.p82-86

Abstract

Background: Candida albicans was found to be dominant in patients with human immunodeficiency virus / acquired immunodeficiency syndrome (HIV/AIDS). The antifungals fluconazole, ketoconazole, and nystatin were used as oral candidiasis therapy for HIV/AIDS, each of which has differing susceptibility in oral candidiasis therapy. Purpose: The present study aimed to evaluate the susceptibility and antifungal resistance to oral C. albicans in HIV/AIDS patients. Methods: The subjects followed the universal precaution principles. Oral Candida species were isolated from the saliva of 98 HIV/AIDS subjects. Identification of Candida species was carried out by the mycobiotic agar of API 20 C Aux system. Susceptibility and resistance antifungal tests on the Candida species were performed using a Fungus ATB Kit. Results: Candida albicans was the most dominant species found from 98 subjects (95%). The rest were other Candida species. There are 41 subjects (42%) with a history of oral candidiasis, and 57 subjects (58%) without. The history of those who used antifungals were: nystatin = 60 subjects (61%), fluconazole = 39 subjects (40%), and ketoconazole = two subjects (2%). These antifungals have a susceptibility above 80% against C. albicans, except the nystatin group (79%) (p>0.05; 0.628), but fluconazole has a strong correlation (r=0.820) to susceptibility, susceptibility-dependent dose, and resistance. Conclusion: Candida albicans was dominant in the saliva of HIV/AIDS patients. This fungus was effectively treated by fluconazole, ketoconazole and nystatin. These antifungals had a high susceptibility at ≤ 8 μg/mL to C. albicans.
Microscopic Examination of Urine Samples as the Early Detection of Asymptomatic Urinary Tract Infection in Pregnant Women: A Cross-Sectional Study: Uji Mikroskopik Spesimen Urin sebagai Deteksi Dini Infeksi Saluran Kemih tidak Bergejala pada Perempuan Hamil: Sebuah Studi Potong Lintang Yeva Rosana; Dwiana Ocviyanti; Rahmah Amran
Indonesian Journal of Obstetrics and Gynecology Volume 7 No. 3 July 2019
Publisher : Indonesian Socety of Obstetrics and Gynecology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.937 KB) | DOI: 10.32771/inajog.v7i3.899

Abstract

Abstract Objectives:To investigate the role of microscopic examination of urine sample in supporting early diagnosis of asymptomatic urinary tract infection (UTI) in pregnant women. To compare correspondence between microscopic examination and urine culture result as a gold standard diagnostic modality to support the diagnosis of UTIs and as an evidence-based to start empirical therapy. Methods: Microscopic analysis was conducted in 74 centrifuged and non-centrifuged urine samples from 317 pregnant who came to six healthcare centres in Jakarta, which showed a positive result of nitrite examination. The results of the microscopic examination of bacteriuria and leukocyturia were compared with a urine culture. Results: Sensitivity of centrifuged bacteriuria was the highest among the other microscopic parameters, which was 74% with the p-value of 0.009. Combination of bacteriuria and leukocyturia≥3/HPF dan≥5/HPF have increased the specificity with the value of 91.5% and 93.6% in non-centrifuged urine. Conclusion: This result showed that the best method of microscopic examination for early diagnosis of asymptomatic urinary tract infection in pregnant women is the detection of bacteriuria in centrifuged urine. Combination of bacteriuriaandleukocyturiatest, as well as leukocyturia≥3/HPF and≥5/HPF, can be used to rule out the diagnosis of UTI at an early stage. Keywords: asymptomatic urinary tract infection, bacteriuria, leukocyturia,microscopic examination, urine culture. Abstrak Tujuan: Mengetahui peran pemeriksaan mikroskopis sampel urin dalam mendukung diagnosis dini infeksi saluran kemih asimptomatik (ISK) pada perempuan hamil. Membandingkan kesesuaian antara pemeriksaan mikroskopis dan hasil kultur urin sebagai modalitas diagnostik standar emas untuk mendukung diagnosis ISK dan sebagai dasar bukti untuk memulai terapi empiris. Metode: Penelitian uji mikroskopik dilakukan pada 74 sampel urin disentrifugasi dan tidak sentrifugasi, dari 317 sampel urin perempuan hamil yang berobat ke-enam puskesmas di Jakarta dengan uji nitritpositif. Hasil uji mikroskopik bakteri uria dan leukosit uria dibandingkan dengan hasil kultururin. Hasil: Sensitivitas bakteriuria yang disentrifugasi menunjukan hasil yang paling baik dibandingkan dengan parameter uji mikroskopik lain, yaitu 74% dengan nilai p yang bermakna sebesar 0,009. Kombinasi bakteri uria dan leukosituria ≥3/LPB dan ≥5/LPB dapat meningkatkan spesifisitas uji dengan nilai 91,5% dan 93,6% pada urin yang tidak disentrifugasi. Kesimpulan: Hasil menunjukkan bahwa bakteri uria pada urin yang disentrifugasi, merupakan metode yang paling baik untuk membantu diagnosis dini ISK tidak bergejala pada perempuan hamil. Uji kombinasi bakteri uria dan leukosituria, serta uji leukosituria ≥3/LPB dan ≥5/LPB dapat dimanfaatkan untuk membantu secara dini menyingkirkan orang yang tidak mengalami ISK. Kata kunci: bakteriuria, infeksi saluran kemih tidak bergejala, kultur urin,leukosituria, uji mikroskopik
Modified Slide Culture Method for Faster and Easier Identification of Dermatophytes YEVA ROSANA; TETSUHIRO MATSUZAWA; TOHRU GONOI; ANIS KARUNIAWATI
Microbiology Indonesia Vol. 8 No. 3 (2014): September 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1069.613 KB) | DOI: 10.5454/mi.8.3.7

Abstract

Basic slide culture as a morphological identification was known as the most common method for the identification of pathogenic mold fungi. This method preserved the morphological features relatively undisturbed compared with adhesive tape preparations. However, it was necessary to modify method of basic slide culture to improve its usability and shorten the time it needed to identify mold fungi. There were four kinds of method carried out in this study; two kinds of modified slide culture, one kind of direct culture on slant agar plate, and a basic slide culture for identifying mold fungi, which result would be compared with each other. These four methods were tested to 4 species of dermatophytes which were known as mold fungi that could infect skin, hair, and nails in human; those were Trichophyton mentagrophytes, Microsporum canis, Microsporum gypseum, and Epidermophyton floccosum. Result of this study showed that both modified slide culture and direct culture on slant agar plate could visualize the structure of dermatophytes faster than basic slide culture method. These methods were also easier to prepare compared to basic culture method. Conclusion of this study showed that basic slide culture method needed to be modified for better identification of mold fungi.
Comparison of Microbial Pattern Causing Urinary Tract Infection in Female Out- and Hospitalized Patients in Jakarta YEVA ROSANA; DWIANA OCVIYANTI; ANIS KARUNIAWATI; SYADZA RHIZKY PUTRI AKHMAD
Microbiology Indonesia Vol. 10 No. 1 (2016): March 2016
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (493.251 KB) | DOI: 10.5454/mi.10.1.5

Abstract

Urinary Tract Infection (UTI) is an infection in any part of the urinary system. Women are 3 times more likely to have UTI than men. The UTI accounts for 15% infection cases in outpatients and 24% cases in hospitalized patients. Although the most common cause of UTI is certain bacteria, but it was not easy to choose the appropriate antimicrobial therapy. Strategy for choosing empiric antimicrobial treatments for UTI in female out- and hospitalized patients should be based on the pattern of the causative organisms. The aim of this study was to understand the microbial pattern causing UTI in female out- and hospitalized patients in Jakarta. The UTI -1 causative microorganisms were obtained from urine culture containing 100,000 cfu/mL . Twenty nine microorganisms were found as the causative agents of UTI in 317 pregnant women who came to six Community Health Centres (Puskesmas) in Jakarta: Makassar; Pulogadung, Cakung, Pasar Rebo, Duren Sawit, and Kramat Jati for antenatal care. Twenty nine microorganisms were isolated from 114 urine samples of female hospitalized patients who were diagnosed of UTI. The samples were obtained from the Microbiology Laboratory Clinic of FKUI-RSCM. The most common microorganisms causing UTI in female out- and hospitalized patients were Gram negative bacteria. In female outpatients, Klebsiella sp was the most common causative bacteria (31%), followed by Escherichia coli (24.1%). In female hospitalized patients, Escherichia coli was the most common causative bacteria (30%), followed by Candida sp (24.1%) and Klebsiella pneumonia (6.8%). There was more variation in the pattern of UTI causative organisms in hospitalized female patients in comparison to that of the outpatients. Candida sp. was only found in hospitalized UTI patients but not in outpatients.
Molecular Viability Assay: Improving Leprosy Diagnosis beyond Current Gold Standard Imaniar, Clara; Ibnu Agus Ariyanto; Yeva Rosana
EKSAKTA: Berkala Ilmiah Bidang MIPA Vol. 26 No. 04 (2025): Eksakta : Berkala Ilmiah Bidang MIPA (E-ISSN : 2549-7464) In Progress
Publisher : Faculty of Mathematics and Natural Sciences (FMIPA), Universitas Negeri Padang, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24036/eksakta/vol26-iss04/568

Abstract

Neglected tropical diseases are still part of the health problems faced by the world. One of the neglected tropical diseases that has not yet reached 100% elimination is leprosy. Mycobacterium leprae is the pathogen responsible for leprosy, a chronic infectious disease that affects the skin and peripheral nerves and can lead to significant disability if left untreated. Currently, the gold standard for diagnosis is detecting acid-fast bacilli (AFB) with Ziehl-Neelsen staining; however, this method cannot distinguish between living and dead bacteria, complicating treatment assessment, relapse detection, and resistance tracking. Therefore, more accurate diagnostic instruments that can differentiate bacterial viability are needed. Since M.leprae cannot be cultured in artificial media, molecular-based assays are promising tools for rapid diagnosis. This study aims to identify recent assays for assessing bacterial viability in leprosy. Articles used are limited to the publication year between 2019 until 2024 from databases such as PubMed, ProQuest, Scopus and Google Scholar, using PRISMA methods. After filtration, from 143 articles we found 5 articles that discussed the viability of leprosy-causing bacteria. The selected studies showed that molecular assays to determine bacterial viability can be used and explored to strengthen the existing gold standard for monitoring treatment of leprosy patients
Risiko Reaktivasi Tuberkulosis Laten pada Terapi COVID-19 Abas, Ghina Mutiara; Sjatha, Fithriyah; Rosana, Yeva
Jurnal Biomedika dan Kesehatan Vol 6 No 3 (2023)
Publisher : Fakultas Kedokteran Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/JBiomedKes.2023.v6.363-372

Abstract

The high mortality rate among COVID-19 patients in the acute respiratory distress syndrome (ARDS) phase led to the administration of immunosuppressive drugs. Corticosteroids could block inflammation caused by cytokine storm, and prevent pneumonia, edema, fibrosis, and ARDS. Even though it was believed to have beneficial effects, corticosteroids can suppress T CD4+ and CD8+ cell-mediated immunity reaction through decreased IFNγ production thus leading to reactivation of latent Tuberculosis (LTBI). Therefore, the usage of corticosteroids in the ARDS phase of COVID-19 patients should be carefully given; pre-screening of LTBI may be done to avoid Tuberculosis reactivation.
Profile of multidrug-resistant bacteria causing urinary tract infections in inpatients and outpatients in Jakarta and Tangerang Rosana, Yeva; Herliyana, Lina; Krisandi, Grady; Anggraini Suwarsono, Erike
Universa Medicina Vol. 42 No. 3 (2023)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2023.v42.303-313

Abstract

Background Urinary tract infections (UTIs) caused by bacteria occupy the second highest rank of common infectious diseases in the world. Empirical use of antibiotics may give rise to multidrug-resistant (MDR) bacteria because of irrational prescription. Choice of antibiotics to treat UTIs is limited because of MDR bacteria. Thus, this study was conducted to investigate the bacterial antibiotic susceptibility patterns in inpatients and outpatients in Jakarta and Tangerang. Methods Bacterial isolates were obtained from midstream urine specimens from 43 inpatients and 43 outpatients with UTIs in Jakarta and Tangerang. Bacteria were isolated on blood and MacConkey agar media using colony count method. Isolate identification and their susceptibility patterns were performed using VITEK2 compact system according to manufacturers’ instructions. Data were analyzed using Chi-square test. Results A total of 89 bacterial isolates consisting of 15 bacterial species were successfully isolated from 86 specimens. Gram-negative bacteria were the most common etiology of UTIs in inpatients and outpatients. MDR bacteria were found in 52 of the 89 isolates. ESBL-producing E. coli was the most common MDR bacteria. ESBL-producing E. coli and other MDR bacteria showed good susceptibility to ertapenem, meropenem, amikacin, and tigecycline. There were no significant differences regarding the MDR bacterial count in inpatients and outpatients (p=0.521). Conclusion E. coli was found to be the most common MDR bacteria causing UTIs in inpatients and outpatients in Jakarta and Tangerang. Higher resistance to many antibiotics was found in MDR bacterial isolates in inpatients compared to outpatients. MDR bacteria in outpatient UTIs were highly resistant to commonly used antibiotics.