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Merdekawati, Fusvita
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Biochemistry, Genetics & Molecular Biology Chemistry Dentistry Education Environmental Science Health Professions Immunology & microbiology Mathematics Medicine & Pharmacology Nursing Physics Public Health Social Sciences Veterinary

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Optimasi suhu amplifikasi DNA pada quantitative polymerase chain reaction untuk identifikasi Mycobcterium tuberculosis resistan isoniazid Endarwati, Dwi Veni; Indra, Asep Iin Nur; Hardiana, Acep Tantan; Abror, Yogi Khoirul; Nurhayati, Betty; Merdekawati, Fusvita
Current Biomedicine Vol. 2 No. 2 (2024): July
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.2.2.61-70

Abstract

Background: Tuberculosis (TB) is a disease caused by Mycobacterium tuberculosis and is a serious threat to global health. The methods can be used to detect and identify the bacteria is quantitative polymerase chain reaction (qPCR). In this method, denaturation and extension temperatures are determining factors of success that needs to be optimized. Objective: This study aims to optimize denaturation and extension temperatures in M. tuberculosis DNA amplification. Methods: The research used quasi-experimental design. The denaturation temperature optimized were 93, 94, 95, 96, and 97°C, and the extension temperature optimized were 58, 59, 60, 61, and 62°C. The test sample was a 1 ml sputum sample isolated from a patient with isoniazid-resistant M. tuberculosis. Optimization was performed using seven test primers, namely S315T, S315N, S315I, S315R, S315G, S315L, and R463B with the katG gene target and data analysis using Ms Excel. Data optimization results were processed with Excel by taking the lowest Ct value. Results: The results showed that the optimization temperatures for denaturation were different for each primer used. Primers S315T, S315R, and S315G, optimal with denaturation temperature of 96°C, primer S315N optimal with 94°C, primers S315I and R463B optimal with 93°C, and for primer S315L optimal with 95°C, with the most widely used temperature is 96°C. The optimal extension temperature was 58°C for primers S315T, S315N, S315I, and R463B, at 60°C for primers S315R and S315G, and at 61°C for primer S315L. Conclusion: The optimal denaturation temperature in this study was 96°C and the optimal extension temperature was 58°C.
Konsentrasi dan kemurnian ekstraksi DNA metode sonikasi dan spin column dari sampel dahak penderita tuberkulosis Saputra, Fitrianingsih; Indra, Asep Iin Nur; Djuminar, Ai; Merdekawati, Fusvita; Nurhayati, Betty
Current Biomedicine Vol. 2 No. 2 (2024): July
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.2.2.84-92

Abstract

Background: The Polymerase Chain Reaction (PCR) method can identify Mycobacterium tuberculosis in a sputum sample of a patient with TB (TB). One crucial step to ensure accurate PCR results is the DNA extraction process. Objective: The research aims to compare the concentration and purity of DNA from the sputum of TB patients using ultrasound and spin column extraction techniques. Methods: The research uses descriptive study designs with post-only design strategies. The primary data was derived from 18 sputum specimens from TB patients. Concentration measurement and DNA purity testing using a nanodrop spectroscopic photometer. Results: DNA extraction by ultrasound method has an average concentration of 18.9 ± 8.5 ng/L, with a peak of 37.6 ng/ L. The spin column method produces an average of 55.5 ± 27.9 ng/μL; the peak is 105.0 ng/ μL. The purity value of the DNA extract is in the range of 1.8 ± 2.0 with the ultrasound method of 61% and the spin column of 78%. Conclusion: The sonication method has a lower average concentration and a higher percentage of purity than the spin column method, and there are differences in concentrations and purity values between the two methods.
ANALISIS KADAR INTERFERON GAMMA MENGGUNAKAN METODE FLUORESCENCE IMMUNOASSAY (FIA) DAN ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) PADA PASIEN TUBERKULOSIS Trimulyani, Annisa; Khoirul Abror, Yogi; Marliana, Nina; Merdekawati, Fusvita
Jurnal Kesehatan Siliwangi Vol. 4 No. 3 (2024): JURNAL KESEHATAN SILIWANGI
Publisher : Politeknik Kesehatan Kemenkes Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Tuberkulosis merupakan 1 diantara 10 penyebab kematian tertinggi diseluruh dunia. Berdasarkan data Global Tuberculosis Report 2022, Indonesia berada pada posisi kedua pada tahun 2021 dengan jumlah penderita tuberkulosis terbanyak di dunia. Tuberkulosis disebabkan oleh Mycobacterium tuberculosis yang dapat menyebar melalui udara dari droplet. Infeksi M. tuberculosis dapat dideteksi melalui pemeriksaan imunoserologis, yaitu pemeriksaan Interferon Gamma Release Assays (IGRA). Pemeriksaan IGRA digunakan untuk mengukur produksi sitokin interferon gamma sebagai respon spesifik terhadap antigen M. tuberculosis Early Secretory Antigenic Target-6 (ESAT-6) dan Culture Filtrate Protein-10 (CFP-10) secara in vitro selama 16 - 24 jam. Semakin berkembangnya teknologi saat ini terdapat metode baru yaitu Fluorescence Immunoassay (FIA) yang hasilnya lebih cepat keluar dibandingkan dengan metode Enzyme-Linked Immunosorbent Assay (ELISA) yang sudah direkomendasikan oleh WHO, memerlukan waktu 3 jam sampai hasil selesai. Penelitian ini bertujuan untuk mengetahui ada atau tidaknya perbedaan kadar interferon gamma menggunakan metode Fluorescence Immunoassay (FIA) dan Enzyme-Linked Immunosorbent Assay (ELISA) pada pasien tuberkulosis. Jenis penelitian yang digunakan adalah eksperimen semu dengan desain penelitian static group comparation. Jumlah sampel yang digunakan dalam penelitian sebanyak 30. Hasil kadar interferon gamma menggunakan metode Fluorescence Immunoassay (FIA) didapatkan rata-rata 2.83 IU/mL dan metode Enzyme-Linked Immunosorbent Assay (ELISA) 2.78 IU/mL. Kemudian data yang diperoleh dianalisis menggunakan uji Wilcoxon didapatkan hasil nilai Asymp. Sig. 0.109 > 0.05. Sehingga dapat disimpulkan bahwa tidak terdapat perbedaan yang signifikan kadar interferon gamma menggunakan metode Fluorescence Immunoassay (FIA) dan Enzyme-Linked Immunosorbent Assay (ELISA).
Comparison of Purity and Concentration Values of Mycobacterium tuberculosis DNA Extraction Result from the Boiling and Spin Column Method Febriyanti, Intan; Djuminar, Ai; Merdekawati, Fusvita; Indra , Asep Iin Nur
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 5 No 2 (2023): Combatting Bacterial and Fungal Infections: The Critical Role of Advanced Researc
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v5i2.4771

Abstract

The Polymerase Chain Reaction (PCR) technique is extensively employed in molecular biology to precisely detect Mycobacterium tuberculosis. Prior to conducting PCR, extracting of high-quality genomic Deoxyribonucleic Acid (DNA) is crucial to ensure accurate and reliable results. The primary objective of this study is to conduct a comparative analysis of the purity and concentration of M. tuberculosis DNA acquired through the utilization of the boiling method and the spin column extraction methods. A descriptive comparative research design was employed, utilizing a sample of 16 sputum specimens that had previously been confirmed as positive for M. tuberculosis through Acid-Fast Bacteria (AFB) examination and Molecular Rapid Test (MRT). The extraction of DNA was carried out using the boiling method and the spin column method. Subsequently, the concentration and purity of the extracted DNA were assessed using the NanoDrop Spectrophotometer, and the results were compared. The obtained yield of M. tuberculosis DNA isolates through the boiling method ranged from 9.6 ng/µL to 1258.7 ng/µL, with an average purity value of 1.23. Conversely, for the spin column method, the concentration of M. tuberculosis DNA isolates ranged from 8.7 ng/µL to 207.8 ng/µL, with an average purity value of 1.83. In conclusion, there is a significant difference between the purity and concentration of M. tuberculosis DNA extraction results using the boiling method and spin column methods.
Analysis of Purity and Concentration Escherichia coli DNA by Boiling Method Isolation with Addition of Proteinase-K and RNase Lesiani, Bunga Rossa; Abror, Yogi Khoirul; Merdekawati, Fusvita; Djuminar, Ai
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 5 No 2 (2023): Combatting Bacterial and Fungal Infections: The Critical Role of Advanced Researc
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v5i2.4773

Abstract

Escherichia coli is a leading cause of Urinary Tract Infections (UTIs) in Indonesia, with approximately 180,000 cases reported annually. The more cases of UTIs, the more PCR diagnosis is needed with an accurate, fast, simple, and economical DNA isolation method. However, currently, there is no DNA purification stage from protein and RNA contaminants in the boiling DNA isolation method. This study aimed to investigate the impact of incorporating Proteinase-K and RNase into the boiling DNA isolation method on the purity and concentration of E. coli’s DNA during isolation. The boiling method involved heating to 95°C – 100°C bring to cell lysis and release of cellular components, including DNA. Urine samples were artificially contaminated with E. coli at different McFarland standards (0.25, 0.5, and 1). The boiling DNA isolation method was then performed and then analyzed for purity and concentration using a NanoDrop spectrophotometer. This study demonstrated a positive correlation between Proteinase-K and RNase concentrations used in the boiling DNA isolation method and the subsequent increase in DNA purity and concentration. An increase in DNA purity and concentration was obtained even though it was not statistically significant compared to that without Proteinase-K and RNase addition, with p-values of 0.245 for DNA purity and 0.353 for DNA concentration. Further research is recommended with higher Proteinase-K and RNase concentrations in the boiling DNA isolation method to achieve improved purity and concentration of E. coli DNA. Such enhancements could improve PCR amplification and help diagnose E. coli-related UTIs.
Pengaruh Lama Demam terhadap Positivitas Rate IgM Anti Salmonella typhi pada Pasien Tersangka Demam Tifoid Metode Inhibition Binding Immunoassay Nurhayati, Nuli; Rohayati, Rohayati; Merdekawati, Fusvita; Marliana, Nina
JPP JURNAL KESEHATAN POLTEKKES PALEMBANG Vol 18 No 2 (2023): JPP (Jurnal Kesehatan Poltekkes Palembang)
Publisher : Poltekkes Kemenkes Palembang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36086/jpp.v18i2.1972

Abstract

Latar Belakang: Demam tifoid merupakan suatu penyakit infeksi sistemik yang disebabkan oleh bakteri Salmonella typhi. Manifestasi klinis demam tifoid yang pasti adalah demam. Demam terjadi pada saat tubuh sedang berusaha untuk melawan infeksi patogen dengan aktivasi sistem imun. Respon imun yang khas dimulai dengan peningkatan antibodi IgM terhadap antigen yang menstimulasi (imunogen). IgM mulai terbentuk pada hari ke-3, dan titernya meningkat hingga mencapai puncaknya pada hari ke-14. Pemeriksaan IgM anti Salmonella menggunakan metode IMBI karena memiliki tes spesifisitas dan sensitifitas yang tinggi. Tujuan penelitian ini untuk mengetahui pengaruh lama demam pada pasien demam tifoid terhadap positivitas rate IgM anti Salmonella. Metode: Penelitian dilakukan di Rumah Sakit Kabupaten Sumedang dan Rumah Sakit Kota Bandung, dengan jenis penelitian observasional analitik cross sectional. Pada periode Mei 2023 sampel yang diambil adalah 32 pasien demam tifoid dengan kadar IgM yang diperiksa menggunakan metode IMBI. Pengambilan sampel dilakukan secara purposive sampling dan diolah dengan menggunakan Microsoft Excel. Hasil: Data yang diperoleh dari hasil perhitungan yaitu positivitas rate IgM anti Salmonella typhi pada lama demam 2 hari adalah 0%; 4 hari adalah 25%; 6 hari adalah 75%; 8 hari adalah 50%. Kesimpulan: Hasil penelitian ini menunjukan bahwa lama demam hari ke 2, 4, 6, dan 8 berpengaruh secara terhadap positivitas rate IgM anti Salmonella typhi.
UPAYA PREVENTIF PENYEBARAN TUBERKULOSIS MELALUI PEMBERDAYAAN MASYARAKAT DALAM MENGOLAH DAUN KELOR SEBAGAI SPRAY DESINFEKTAN DI RT 02 RW 02 KELURAHAN PASIRKALIKI, KECAMATAN CIMAHI UTARA, KOTA CIMAHI Abror, Yogi Khoirul; Merdekawati, Fusvita; Juliastuti, Aditya
Jurnal Pengabdian Masyarakat Kesehatan Indonesia Vol. 3 No. 2 (2024): Jurnal Pengabdian Masyarakat Kesehatan Indonesia
Publisher : Poltekkes Kemenkes Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.34011/jpmki.v3i2.2619

Abstract

Tuberculosis (TB) remains a global health issue, with Indonesia ranking second worldwide in TB cases. The high incidence of TB is partly due to its ease of transmission via droplets in densely populated areas. One preventive measure is improving environmental hygiene through disinfectants. However, chemical-based disinfectants often leave residues, prompting the need for natural alternatives, such as Moringa oleifera leaves, which exhibit antibacterial potential. This community service activity aimed to enhance the knowledge and skills of Posyandu Mawar 01 and 02 cadres in RT 02 RW 02, Pasirkaliki Village, Cimahi Utara Sub-district, on processing moringa leaves into disinfectant sprays to prevent TB transmission. The program included preparation, education, mentoring, pre-tests, and post-tests. Supporting materials such as presentations, leaflets, videos, and moringa-based disinfectant spray products were distributed to cadres for further education. The results showed a 42% increase in cadres' knowledge, with an average post-test score of 85 compared to a pre-test score of 60. This activity effectively improved cadres' understanding of TB prevention using natural materials. It is hoped that similar activities will be conducted regularly to raise community awareness and capacity in TB control.
In silico prediction of multi-epitope vaccine candidates against Mycobacterium leprae Shabrina, Almas; Indra, Asep Iin Nur; Rinaldi, Sonny Feisal; Merdekawati, Fusvita
Current Biomedicine Vol. 3 No. 1 (2025): January
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.3.1.22

Abstract

Background Leprosy, also known as Hansen's disease, is an infectious disease caused by Mycobacterium leprae. Despite ongoing efforts to control the disease, leprosy remains a global health concern, with Indonesia ranking third in the world for the highest number of cases. Objective This study aims to identify epitopes that can induce T and B cell immune responses through an in silico approach, to design a multi-epitope vaccine candidate against Mycobacterium leprae. Methods The study used an in silico vaccine design approach utilizing ESAT6, Ag85B, ML2028, ML2380, and ML2055 proteins from Mycobacterium leprae. The process involved sequence alignment, T cell (CTL and HTL) and B cell epitopes identification, and antigenicity, allergenicity, and toxicity assessment. Selected epitopes were constructed into a multi-epitope vaccine candidate using linkers. The tertiary structure of the vaccine was modeled with AlphaFold and evaluated via Prosa-web. The stability and interaction between the vaccine candidate and TLR4 were analyzed using molecular docking. Results The vaccine candidate demonstrated stable interactions with TLR4, with a binding free energy of -13.9 kcal/mol. The vaccine candidate was also predicted to be stable, antigenic, non-allergenic, non-toxic, and hydrophilic. Conclusion This in silico design of a multi-epitope vaccine candidate shows potential for development as a vaccine against leprosy.
Correlation of polymerase chain reaction results with hematocrit levels and platelet counts in dengue patients in Batam City Simangunsong, Kristina; Nurhayati, Betty; Hayati, Eem; Merdekawati, Fusvita
Current Biomedicine Vol. 3 No. 1 (2025): January
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.3.1.1

Abstract

Background Dengue hemorrhagic fever (DHF) is a viral disease transmitted by Aedes aegypti mosquitoes, posing global public health challenge. The Riau Islands Province has the highest incidence of DHF in Indonesia. Objective This study aimed to investigate the relationship between hematocrit and platelet levels with the cycle threshold (Ct) values of polymerase chain reaction (PCR) results in DHF cases in Batam City, Riau Islands Province. Methods A descriptive correlation study was conducted using data from 102 patients infected with the dengue virus. Hematocrit and platelet counts were measured using a hematology analyzer, while Ct values for DENV1, DENV2, DENV3, and DENV4 were obtained through real-time qRT-PCR. Pearson's correlation test was employed to analyze the relationship between these variables. Results The study found no significant gender difference in DHF incidence (males: 50%, females: 50%). The highest prevalence was observed in the 6–11 years age group (44.1%), followed by the 12–18 years group (25.5%), the >18 years group (24.5%), and the 1–5 years group (11.8%). DENV3 was identified as the dominant serotype. No statistically significant correlation was found between Ct values and hematocrit (p = 0.607) or platelet counts (p = 0.323). Conclusion DHF cases in this study showed no gender disparity, with the most affected group being children aged 6–11 years, and DENV3 was the prevalent serotype. Ct values did not show a statistically significant correlation with hematocrit levels or platelet counts, suggesting that these hematological parameters may not predict viral load in DHF cases.
Sensitivity and specificity of the lipoarabinomannan test compared to GeneXpert in urine samples for tuberculosis diagnosis Irawan, Danni; Rismiarti, Zuri; Tantan, Acep; Merdekawati, Fusvita
Current Biomedicine Vol. 3 No. 1 (2025): January
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.3.1.7

Abstract

Background Mycobacterium tuberculosis (MTB) is the causative agent of tuberculosis (TB), primarily affecting lung tissue but also capable of infecting pleura, lymph nodes, bones, and other extrapulmonary sites. Lipoarabinomannan (LAM) is a critical lipopolysaccharide in the outer wall of mycobacterial cells and can be detected in the urine of TB patients as an antigen. Objective This study aimed to assess the sensitivity and specificity of the LAM test compared to GeneXpert in urine samples from suspected TB patients. Methods A quasi-experimental design was employed, where urine samples were collected from patients diagnosed with TB at Sidawangi Lung Hospital, West Java Province. The LAM test was performed on 40 samples by applying 60 µL of urine onto LAM test strips, while MTB presence in urine was examined using GeneXpert. Results LAM test results showed 32.5% positivity, while 67.5% were negative. GeneXpert results indicated 20% positivity and 80% negativity. The LAM test demonstrated a sensitivity of 100% and specificity of 79.4% compared to GeneXpert, with an area under the curve (AUC) value of 0.897. Conclusion The LAM test showed high sensitivity and moderate specificity compared to GeneXpert in urine samples of suspected TB patients.
Co-Authors Abror, Yogi Khoirul Adrian Prasetya, Rifky Dewi Nurhayati Djuminar, Ai Dzulhizza, Zaskia Puteri Endarwati, Dwi Veni Endrique Putri, Salmanda Ernawati Ernawati Febriyanti, Intan Hardiana, Acep Tantan Hayati, Eem Heri Setiyo Bekti I Gusti Agung Ayu Dharmawati Iin Nur Indra, Asep Iis Kurniati Indra , Asep Iin Nur Indra, Asep Iin Nur Irawan, Danni Juliastuti, Aditya Khairunnisa, Syirin Nisrina Khoirul Abror, Yogi Kurnaeni, Nani Lesiani, Bunga Rossa Nina Marliana, Nina Noviar, Ganjar nur amani putri, afifah Nur Habibah Nurhayati, Betty Nurhayati, Nuli Pratama, Resta Ratna Juwita Rahmawati, Tia Rinaldi, Sonny Faisal Rinaldi, Sonny Feisal rohayati rohayati Saputra, Fitrianingsih Shabrina, Almas Simangunsong, Kristina Solihat, M. Firman Sufa, Hafizah Ilmi Suryawan, Muhammad Raihan Syafitri, Andita Izmi Tantan, Acep Trimulyani, Annisa Zanuba, Haifanisya Zahra Zuri Rismiarti