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All Journal HAYATI Journal of Biosciences ACTA VETERINARIA INDONESIANA Jurnal Penelitian Pendidikan IPA (JPPIPA) Makara Journal of Science Jurnal Kedokteran Hewan
UUS SAEPULOH
PRC IPB
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Education Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Physics Veterinary

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Development and Optimization of an Immunoassay for the Detection of Antibodies Against SARS-CoV-2 with In-house Recombinant RBD Protein Ratu, Safira Pinaka Pramestika; Mariya, Silmi; Noviana, Rachmitasari; Saepuloh, Uus; Darusman, Huda Salahudin
Makara Journal of Science Vol. 26, No. 3
Publisher : UI Scholars Hub

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

Abstract

COVID-19 caused by SARS-CoV-2 poses a major threat to the global community, particularly in Indonesia. Countermeasures to prevent the spread of this disease have also been implemented, including the implementation of a vaccination program. An immunoassay technique that can be used to analyze antibodies that might develop following vaccination is the indirect enzyme-linked immunosorbent assay (ELISA). We produced the recombinant spike protein used in this study. The optimization comprised adjusted concentrations of spike recombinant protein (5 and 10 ng/mL), blocking agent (2.5% and 5%), and conjugate (1:1000 and 1:5000). The optimal conditions in this study included a spiked concentration of 10 ng/mL, a blocking agent concentration of 5%, sample dilution of 1:33, and a conjugate concentration of 1:1000. The intra-assay value of this optimized indirect ELISA was 7.3, and the inter-assay value was 5.3. The commercial MyBioSource kit and immunodiagnostic were utilized as a reference in the T-test, with P-values of 0 and 0.313, indicating that the recombinant protein in-house ELISA kit in this study demonstrated the same ability as the commercial immunodiagnostic kit in detecting SARS-CoV-2 antibodies, allowing it to be used for post-vaccination efficacy evaluation.
Genetic Variability of the Long-Tailed Macaque (Macaca fascicularis) Populations in Urban Habitat in Padang City, West Sumatra, Indonesia Sari, Ruhama Maya; Saepuloh, Uus; Rizaldi; Perwitasari-Farajallah, Dyah
HAYATI Journal of Biosciences Vol. 31 No. 2 (2024): March 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.2.392-403

Abstract

The long-tailed macaque (Macaca fascicularis) is a primate species recognized for its exceptional ability to adapt to urban habitat. However, urban anthropogenic activities contribute to the fragmentation of macaque natural habitat, affecting genetic variation among distinct populations. Therefore, this study aimed to assess the genetic variability of M. fascicularis populations in Padang City, West Sumatra, Indonesia. A total of 70 fecal samples from the wild long-tailed macaques in Gunung Padang (GPD), Gunung Meru (GMR), and Gunung Pangilun (GPG) were collected using a non-invasive method. Conventional PCR amplification and Sanger sequencing were conducted to examine a 1,200-bp mitochondrial DNA (mtDNA) fragment in the D-loop region. The analysis of genetic variation showed that only two haplotypes were present in the three populations. Both GPD and GMR shared the same haplotype (H1), while the GPG population had a distinct haplotype (H2). No intrapopulation variation was observed, and haplotype differences were found in ten nucleotide sites with transition substitution mutations. These results showed limited genetic variation among populations of the long-tailed macaque in Padang, thereby providing valuable insights for stakeholders when formulating genetic conservation policies.
RELIABILITY OF AMYLOID DETECTION IMMUNOASSAY KITS FOR ALZHEIMERS DISEASE SCREENING: A PRELIMINARY STUDY Putri, Indah Aprianti; Ningrum, Emilna Mega; Noviana, Rachmitasari; Mariya, Silmi; Saepuloh, Uus; Retnani, Elok Budi; Hamdan, Muhammad; Nugraha, Jusak; Darusman, Huda Shalahudin
Jurnal Kedokteran Hewan Vol 18, No 3 (2024): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v18i3.31109

Abstract

Alzheimers Disease is a progressive neurodegenerative disease that damages the brain and is part of dementia. Alzheimers dementia is caused by various factors that until now the proper therapy is uncertain. Kit production as a diagnostic tool to support the development of screening for Alzheimers disease is a strategic effort. Screening based on detection with Enzyme-Linked Immunosorbent Assay (ELISA) can work effectively and accurately to optimize a specific antibody material or monoclonal antibody to the A42 peptide. This study aimed to research the reliability of the amyloid detection kit which was developed as an alternative screening test for Alzheimers disease compared to commercially available kits. Based on the results and interpretation of inter and intra-assay coefficients, the in-house ELISA kit has comparable A42 detection to the commercially available ELISA kit
Identification of Herpesviridae in Macaca Fascicularis Using the Nested PCR Method at PSSP IPB Malik, Abdul; Sulistiawati, Erni; Yuliana; Saepuloh, Uus
Acta VETERINARIA Indonesiana Vol. 13 No. 1 (2025): Maret 2025
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/avi.13.1.60-66

Abstract

PCR technique is used to detect the presence of Herpesviridae viruses in Macaca fascicularis that exhibit no clinical symptoms and identify them using nucleotide base sequences. This study aimed to detect and identify the presence of nucleotide base sequences in the Herpesviridae virus family in Macaca fascicularis at PSSP IPB. The study utilized 20 nasal swab samples from Macaca fascicularis. Extraction of DNA utilizing the QiaAmp™ DNA mini kit and subsequent measurement of DNA concentration. Amplification of target DNA utilizing the nested PCR technique. Sample with positive results from electrophoresis were sequenced to obtain nucleotide sequences. The average measurement value of DNA extraction concentration was 25.07 ng/µl. The DNA purity ratios at wavelengths A260/A280 and A260/A230 averaged 1.95 and 0.27, respectively. The electrophoresis results indicated a band size of 215 bp in sample code 01, corresponding to the positive control band. The sequencing results were analyzed using BLASTn on the NCBI site, revealing similarity with Macaca fascicularis lymphocryptovirus. The BLAST sample results demonstrated a Query Cover value of 100% and a percentage of identity of 98.84%.
THE GENE EXPRESSION OF ADAM17 AS A GENETIC MARKER OF ALZHEIMER DISEASES IN THE BRAIN OF LONG-TAILED MACAQUES (Macaca fascicularis) Rosmanah, Lis; Hasya, Karin katina; Saepuloh, Uus; Manalu, Wasmen; Winarto, Adi; Sudarnika, Etih; Darusman, Huda Shalahudin
Jurnal Kedokteran Hewan Vol 17, No 3 (2023): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v17i3.31014

Abstract

This study aims to identify the expression of the a disintegrin and metalloproteinase 17 (ADAM17) gene as a marker of Alzheimer's disease in long-tailed macaques (Macaca fascicularis). This study used six brain samples (hippocampus and cortex regions) of long-tailed macaques which was divided into two groups consisting of aged long-tailed macaques and adult long-tailed macaques. The expression of ADAM17 gene was determined by comparing the relative quantification values between the two age groups, and brain regions consisting of the hippocampus and cortex regions. The results of data analysis showed no significant difference in the expression of ADAM17 gene between brain regions and between age groups of long-tailed macaques. However, numerically the results showed a lower expression of ADAM17 gene in the hippocampus region of aged macaques. Lower expression of ADAM17 gene could be a marker of old animals indicating the stages of Alzheimer's disease.
IDENTIFICATION AND MOLECULAR CHARACTERIZATION OF BOVINE HERPERVIRUSES (BoHV) DNA TERMINASE PARTIAL GENE IN ACEH CATTLE Prayitno, Lilik; Saepuloh, Uus; Mayasari, Ni Luh Putu Ika; Faisal, Faisal; Ayuningsih, Ellis Dwi; Pamungkas, Joko Pamungkas
Jurnal Kedokteran Hewan Vol 11, No 4 (2017): December
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i4.8024

Abstract

Bovine Herpesvirus (BoHV) is a member of Herpesviridae family that acts as pathogenic virus causing infectious bovine rhinotracheitis (IBR) among cattles, resulting in economic loss for cattle industry. BoHV-1 infection in cows is closely related to abortion, respiratory infection, reduced milk production, infertility, and low birth weight. The aim of this study was to identify and characterize the molecular of BoHV-1 and other virus types, as well as the possible presence of other Herpesviridae family using PCR to amplify DNA terminase gene. Four out of 210 nose swab samples were positive for herpes virus on DNA terminase gene. Further characterization of samples showed 99-100% similarity to BoHv-1 and BoHV-6 sequence. Genetic distance between genera BoHV-1 and BoHV-6 is 0.518 and within genera was 0.001 and 0.044. According to phylogenetic tree analysis of DNA terminase gene, the analyzed sequence clustered into 2 genera, namely Varicellovirus which is identical to BoHV-1 and Macavirus which is identical to BoHV-6. The study provides scientific information on molecular characteristics of Herpesviridae family, especially BoHV-1 which is prevalent in Indonesia with the highest density in the central ranches in Aceh province
Development of Biotin and Horseradish Peroxidase Labelling against Monoclonal Antibody Amyloid-β42 and Its Application on Enzyme-Linked Immunosorbent Assay Fiero, Nur Rizky; Saepuloh, Uus; Darusman, Huda Shalahudin; Noviana, Rachmitasari
Jurnal Penelitian Pendidikan IPA Vol 11 No 10 (2025): October
Publisher : Postgraduate, University of Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29303/jppipa.v11i10.12528

Abstract

Alzheimer disease is one of the causes of dementia that could make the patient lose their memories until they unable to do normal activities. It is caused by accumulation of protein in the brain called Amyloid-β42 (Aβ42), that often used as biomarker for alzheimer by enzyme-linked immunosorbent assay (ELISA). To increase the sensitivity and accuracy, capture antibodies are conjugated with labels such as Horseradish Peroxidase (HRP) and Biotin. However, the two conjugates also differ in sensitivity and specificity, therefore this research is to find which is more optimum and stable for ELISA usage. The molecular mass of the conjugated antibodies was characterized with SDS-PAGE, result shows the pre-conjugated antibody has 150 kDa bands, conjugated antibody with labels is heavier than the pre-conjugated due to the addition of each label. Conjugated antibody tested with tetramethylbenzidine (TMB), then analyzed by ELISA reader. The measurement precision decided with %CV value with results shows dillution from 10-1 to 10-4 has %CV less than 10%, while Biotin is from 10-1 to 10-5. Optimization done by determining fixed antigen with the conjugated antibody, the optimal concentration for HRP is 10-3 while biotin is at 10-4. Stability of the conjugated antibody was also determined, HRP measurement started to unstable around second month and third month, while biotin start to unstable around fourth month, this shows the biotin is more stable than the HRP.
Co-Authors Abdul Malik Adi Winarto DYAH PERWITASARI -FARAJALLAH Ellis Dwi Ayuningsih Elok Budi Retnani Erni Sulistiawati Etih Sudarnika Faisal Faisal Fiero, Nur Rizky Hasya, Karin katina Huda Shalahudin Darusman Jusak Nugraha Mayasari, Ni Luh Putu Ika Muhammad Hamdan Ningrum, Emilna Mega Noviana, Rachmitasari Pamungkas, Joko Pamungkas Prayitno, Lilik Putri, Indah Aprianti Ratu, Safira Pinaka Pramestika Rizaldi Rosmanah, Lis Sari, Ruhama Maya SILMI MARIYA Wasmen Manalu Yuliana Yuliana