Research Journal of Pharmacy and Technology

Nelly Zuroidah ¹, May Fanny Tanzilia ², I Gusti Agung Ayu Eka Putri Sunari ¹, Billy Jordan Wrahatnala ³, Faradila Khoirun Nisa Haki ³, Aryati ⁴, Puspa Wardhani ⁵, Dominicus Husada ⁶, Ali Rohman ⁷, Siti Nadia Tarmizi ⁸

Affiliations
1 Clinical Pathology Specialization Program, Faculty of Medicine, Universitas Airlangga, Surabaya, ID
2 Clinical Pathology Sub-Specialization Program, Faculty of Medicine, Universitas Airlangga, Surabaya, ID
3 Master Program of Tropical Medicine, Faculty of Medicine, Universitas Airlangga, Surabaya, ID
4 Department of Clinical Pathology, Faculty of Medicine, Universitas Airlangga, Surabaya, ID
5 Institute of Tropical Disease, Universitas Airlangga, Surabaya, ID
6 6Department of Pediatric, Faculty of Medicine – Universitas Airlangga, Surabaya., ID
7 Department of Chemistry, Faculty of Science and Technology – Universitas Airlangga, Surabaya., ID
8 Directorate of Vector Borne and Zoonotic Disease, Ministry of Health Republic of Indonesia, Jakarta,, ID

Abstract

Background : NS1 is currently widely used for diagnosis of dengue virus (DENV) infection. Various methods are used to diagnose DENV infection (DVI), either ELISA, immunochromatography (ICT) or most recently the fluorescence immunoassay (FIA) method which are commercially available. Objective: This study aimed to compare the detection capabilities of dengue NS1 antigens using (1) Dengue NS1 ICT Ag (Standard Q - SD Biosensor, Inc.), (2) Dengue NS1 ICT Ag (SD Bioline - Standard Diagnostic, Inc), and (3) Dengue NS1 Ag FIA (Standard F - SD Biosensor, Inc.) Methods: This study consisted of serum samples (n=80) with the number of DVI patients (n=50), non-DVI (n=30). All samples were examined using all three commercial kits for NS1 antigen testing. All DVI samples showed results of reverse-transcriptase polymerase chain reaction (RT-PCR - SIMPLEXAᵀᴹ Dengue - Focus Diagnostics) and/or positive dengue NS1 (Panbio® Dengue Early ELISA) antigen. Results: Standard F showed the highest sensitivity (82%) compared to Standard Q (74%) and SD Bio line (74%). These three commercial kits had the same specificity 100%. The positive predictive value all of these kits was 100% each. The negative prediction value of Standard F, Standard Q, and SD Bio line were 76.9%, 63.8%, 63.8%, respectively. These three NS1 antigen tests had a good agreement (κ 0.681-0.774). Conclusions: FIA test performance (Standard F SD - Biosensor, Inc.) were a quick and easy examination, showing a higher sensitivity and specificity than ICT for detecting DENV infection. Further research is needed to confirm the diagnosis of primary or secondary infection.

Keywords

Dengue Viral Infection, NS1 Dengue antigen, ICT, FIA.

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Lukisiari Agustini ¹, Nurwasis ¹, Aryati ², Gatut Suhendro ¹, I Ketut Sudiana ³, Widjiati ⁴, Bambang Purwanto ⁵, Winarto ⁶

Affiliations
1 Department of Ophthalmology, Faculty of Medicine-Dr. Soetomo Teaching Hospital, Universitas Airlangga, Surabaya 60131,, ID
2 Department of Pathology Clinic, Faculty of Medicine-Dr. Soetomo Teaching Hospital, Universitas Airlangga, Surabaya 60131,, ID
3 Department of Anatomic Pathology, Faculty of Medicine-Dr. Soetomo Teaching Hospital, Universitas Airlangga, Surabaya 60131,, ID
4 Department of Veterinary Anatomy, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115,, ID
5 Department of Physiology, Faculty of Medicine-Dr. Soetomo Teaching Hospital, Universitas Airlangga, Surabaya 60131, ID
6 Faculty of Medicine, Universitas Diponegoro, Semarang 50275,, ID

Abstract

Background: The cause of cell death is thought to be due to the pathological apoptotic process in Retinal Ganglion Cells (RGCs), but how the exact mechanism of what is most influential is still not explained. Objective: This study aimed to explain the mechanism of RGCs apoptosis Rattus Norvegicus which is thought to underlie the occurrence of ethambutol toxic optic neuropathy. Methods: A total of 42 male, adult Rattus norvegicus Sprague-Dawley strains were divided into 6 groups with 3 control groups and 3 treatment groups in a randomized design with time series test. The treatment groups were given ethambutol 15 mg/kg/day for each group within 5, 10 and 15 days orally using a gauge.Expressions of SOD2, MDA, PKC δ, p53, Cyt c, Caspase 3 and apoptosis were examined by immunohistochemical methods. Results: Ethambutol affected significant decreased expression of SOD2 with p=0.002 in 5 days, p=0.013 in 10 days and p=0.018 in 15 days; significant increased MDA in 5 days with p=0.05, 10 days with p=0.017, 15 days with p=0.002; significant increased p53 in 5 days with p=0.012, 10 days with p=0.002, 15 days with p=0.001; significant increased Cyt c in 5 days with p=0.004, 10 days with p=0.001, 15 days with p=0.001; significant increased Caspase 3 in 5 days with p=0.001, 10 days with p=0.003, 15 days with p=0.001 and apoptosis in 5 days with p=0.001, 10 days with p=0.001, 15 days with p=0.001. Conclusion: The mechanism of apoptosis of RGCs caused by ethambutol was showed via decreased expression SOD2, increased expression of MDA, p 53, Cyt c, Caspase 3 andapoptosis. These biomarkers are essential to detect apoptosis as one of mechanism in cell death.

Keywords

Apoptosis, ethambutol, toxic, Rattus, mechanism, SOD2.

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