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Journal : INDONESIAN JOURNAL OF MEDICAL LABORATORY SCIENCE AND TECHNOLOGY

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.
ANALYSIS OF APO-B SERUM LEVELS IN BALB/C MICE HYPERCHOLESTEROLEMIC AGAINST TEMULAWAK EXTRACT (Curcuma xanthoriza Roxb) Solihah, Riyadatus; Haris, M. Shofwan; Abror, Yogi Khoirul
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 1 No 1 (2019): The Value, Importance, and Oversight of Health Research
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v1i1.799

Abstract

APO–B Serum levels is the most predictive value for the incidence of atherosclerosis and coronary heart disease. Curcuma xanthorrhiza Roxb contains curcumin, which can be used as an antioxidant, anti–inflammatory and antihypercholesterol. The mechanism of curcumin contained in ginger to reduce cholesterol is due to its function as a cholagoga or bile stimulant. This study aims to determine the effect of temulawak extract on the levels of APO–B Serum in hypercholesterolemia mice. This research were a true experimental study with a post–test only control group design carried out in February 2018. The extraction As much as 25 mice were divided into 5 groups where are group consisted of 5 mice. Positive control group (K+) were treated with high cholesterol feed and water, negative control group (K–) were given standard feed and water, treatment group 1 (P1) were given high cholesterol food and 25mg/kg BW of curcuma extract for 14 days, treatment group 2 (P2) were treated with foods high in cholesterol and 50mg/kg BW of curcuma extract for 14 days and treatment group 3 (P3) treated with high cholesterol and ginger extract 75mg/kg BW for 14 days. Examination of APO–B levels were measured using the spectrophotometric method. Data were analyzed using One–Way Anova. The results showed that the average of APO–B level at (K+) was 209.7 ± 1.02 mg / dL, at (K–) 115.3 ± 1.04 mg / dL, at (P1) 180.4 ± 1.07 mg / dL, at (P2) 147.6 ± 1.12 mg / dL, at (P3) 119.1 ± 1.10 mg / dL. Based on the results of statistical test it was found that there was a significant decrease in APO–B levels with p–value= 0.001 at alpha 0.05 (p <α).