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All Journal Meditory : The Journal of Medical Laboratory Communications in Science and Technology Jurnal Analis Kesehatan Jurnal Riset Kesehatan Poltekkes Depkes Bandung Journal of Indonesian Medical Laboratory and Science (JoIMedLabS) JMLS: Journal of Medical Laboratory and Science Current Biomedicine Mukhtabar: Journal of Medical Laboratory Technology Jurnal Kesehatan Siliwangi Jurnal Pengabdian Masyarakat Kesehatan Indonesia Jurnal Media Analis Kesehatan Poltekita: Jurnal Pengabdian Masyarakat
Nurhayati, Betty
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Biochemistry, Genetics & Molecular Biology Chemistry Dentistry Education Engineering Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology Nursing Public Health Veterinary

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Journal : Current Biomedicine

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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.
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.
Co-Authors A, Nisa Abror, Yogi Khoirul Arbie, Arbie Candra Purnawan Darmantoputro, Feby Wicaksono Dermawan, Asep Didit Damayanti Djuminar, Ai Durachim, Adang Edi Pramono Eka Setiawan, Regina Endarwati, Dwi Veni Ernawati Ernawati Firman Solihat, Mohamad Gusmana, Riki Hardiana, Acep Tantan Hayati, Eem Ida Widiawati, Ida Ihsan Nurhadi, Muhammad Indra, Asep Iin Nur Juliastuti, Aditya Kristian, Erick Kurnia Kusniasih, Susi Lestari, Felia Maryani, Nita Merdekawati, Fusvita Nina Marliana, Nina Noviar, Ganjar Nur Ramadhani, Riski Nurhadi, Muhammad Ihsan Ozi Adi Saputra, Ozi Adi Pratama, Resta Ratna Juwita Pujiasih, Septi Ramadhani, Riski Nur Ridwanna, Surya Rinaldi, Sonny Faisal Rinaldi, Sonny Feisal Rizki, Vanani Nur Rumahorbo, Hotma Saputra, Fitrianingsih Sasmita, Anah Simangunsong, Kristina Suhartini, Entin Suryawan, Muhammad Raihan Tania, Monica Thayyiba, Amina Wardhani, Azkya Zhahira