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Diagnostic Value of Saliva RT-PCR Test within Suspected SARS-CoV-2 Cases in Indonesia Putra, Andika Chandra; Zaini, Jamal; Ridwanuloh, Asep Muhammad; Nugroho, Herjuno Ari; Setyawan, Ryan Haryo; Idris, Idris; Setiawan, Ruby; Sushadi, Pangda Sopha; Wulandari, Ari Sulistyo; Zannati, Anky; Indriawati, Indriawati; Erdayani, Eva; Wahyuni, Wahyuni; Agustiyanti, Dian Fitria; Wisnuwardhani, Popi Hadi; Saniyyah, Zahrah; Azika, Wira Norman; Haryanto, Budi; Utomo, Ahmad Rusdan Handoyo; Ningrum, Ratih Asmana
Health and Medical Journal Vol 6, No 2 (2024): HEME May 2024
Publisher : Universitas Baiturrahmah

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33854/heme.v6i2.1494

Abstract

Introduction: The ongoing SARS-CoV-2 pandemic has profoundly emphasized the pressing need for accurate and reliable diagnostic procedures. Given the potential health risks associated with nasopharyngeal swabs, there has been growing interest in seeking alternative diagnostic mediums. In this context, our study delved into evaluating saliva as a potential diagnostic tool, simultaneously assessing its efficiency in relation to patient demographics and their exhibited clinical symptoms. Methods: Spanning from May to December 2020, we conducted a comprehensive cross-sectional analysis. We meticulously examined medical records to gather insights on patient characteristics, existing health conditions, onset of symptoms, clinical manifestations, and compared the results obtained from both salivary and nasopharyngeal RT-PCR tests for SARS-CoV-2. Results: Among the individuals suspected of SARS-CoV-2 infection, the mean age stood at 52.4 years, with males representing 60.3% of this group. Interestingly, a significant 76.9% reported underlying health conditions, predominantly hypertension and diabetes. The most commonly reported symptoms encompassed respiratory challenges, notably coughing and shortness of breath, succeeded by symptoms like nausea, fever, and a general sense of fatigue. The performance of saliva tests, in terms of accuracy, appeared to be significantly influenced by the timing of symptom emergence. Conclusion: The RT-PCR tests utilizing saliva samples demonstrated considerable promise, especially during the early stages of symptom manifestation, providing a reliable alternative to traditional nasopharyngeal swabs. The findings suggest a superior diagnostic sensitivity when utilizing saliva during the initial phases of a SARS-CoV-2 infection.
Evaluation of Curcumin-derived Carbon-dots' Inhibitory Activity as SARS-CoV-2 Antiviral Candidate Using Chemical Crosslinking Taharuddin, Audrey Angelina Putri; Yamahoki, Nicholas; Stephanie, Rebecca; Agustiyanti, Dian Fitria; Wisnuwardhani, Popi Hadi; Angelina, Marissa; Rubiyana, Yana; Ningrum, Ratih Asmana; Wardiana, Andri; Desriani, Desriani; Hariyatun, Hariyatun; Iskandar, Ferry; Permatasari, Fitri Aulia; Giri-Rachman, Ernawati Arifin; Fibriani, Azzania
HAYATI Journal of Biosciences Vol. 33 No. 1 (2026): January 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.1.232-239

Abstract

In our previous work, we demonstrated that curcumin-derived carbon dots (Cur-CDs) have potential as antivirals for COVID-19. However, the precise mechanism of action remains unclear. This study investigated the potential of Cur-CDs against SARS-CoV-2 by targeting the dimerization of the C-terminal domain of nucleocapsid protein (N-CTD) using chemical crosslinking. Recombinant SARS-CoV-2 N-CTD was expressed, purified, and subjected to chemical crosslinking. The dimerization inhibition ability of Cur-CDs was assessed with ligand concentrations ranging from 0 to 2,000 μg/mL. Successful inhibition —defined as a noticeable reduction in SARS-CoV-2 N-CTD dimer band intensity on SDS-PAGE—was observed when Cur-CDs were present at 8 to 16 times the protein concentration. We hypothesize that Cur-CDs bind to the dimerization residues, preventing non-covalent interactions between monomers and limiting dimer formation. Our findings suggest that Cur-CDs could be a promising antiviral strategy for SARS-CoV-2, especially targeting the dimerization of the nucleocapsid protein. Additionally, this study also highlights the use of chemical crosslinking as a valuable tool for interaction-based drug screening.
Overproduction and Purification of Soluble Recombinant Human Granulocyte Colony Stimulating Factor in Escherichia coli Using Thioredoxin as Fusion Agustiyanti, Dian Fitria; Retnoningrum, Debbie Sofie; Rachmawati, Heni; Fuad, Asrul Muhamad
Annales Bogorienses Vol. 21 No. 1 (2017): Annales Bogorienses
Publisher : BRIN

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

Abstract

Recombinant human Granulocyte Colony Stimulating Factor (G-CSF) has been produced in a soluble form in Escherichia coli BL21 (DE3) as a fusion protein. The open reading frame of G-CSF was synthetically constructed in previous work and was codon optimized for best expression in E. coli. In this research, the gene was fused to thioredoxin (Trx) at the N-terminal in pET32 vector. The purpose of this research was to optimize the overproduction and purification processes to obtain high yield recombinant protein in soluble form, and to characterize the Trx-G-CSF fusion protein. Overproduction was performed using IPTG induction method for 3 and 6 hours. The protein was purified by Ni-NTA affinity chromatography and separated using gradient concentration of imidazole. The purified protein was then characterized by SDS-PAGE and Western Blot analysis. Further, enterokinase was used to separate G-CSF from the fusion protein. The purified form of G-CSF was subsequently characterized using Western Blot and mass spectrometry using MALDI-TOF. The results showed that the fusion protein was successfully produced in soluble part as much as 48.25% were obtained after 3 hours of induction. The yield of fusion protein was 67.37% from total protein (229.65 mg protein/L culture). The Western Blot analysis showed the G-CSF band at around 18.6 kDa. Mass spectrometry with MALDI-TOF/ TOF revealed that 25.86% of amino acid residue was recognized as part of human G-CSF sequence.