Evaluation of NS1-Detection-Based Cell Culture Method for Isolation of Dengue Viruses from Clinical Samples
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MEDICINE
Evaluation of NS1-Detection-Based Cell Culture Method for Isolation of Dengue Viruses from Clinical Samples Shubham Shrivastava 1 & Anamika Solaskar 1 & Mrunal Gosavi 1 & Divya Tiraki 1 & Akhilesh Chandra Mishra 1 & Vidya A. Arankalle 1 Accepted: 1 April 2020 / Published online: 4 May 2020 # Springer Nature Switzerland AG 2020
Abstract Dengue is the fastest spreading mosquito-borne viral infection across the globe affecting over 40% of world populations. Coexistence of 4 distinct serotypes, disease severity due to secondary infection, and continuous evolution of dengue viruses over the years intensify the need for a better and effective virus surveillance program. In this study, we evaluated NS1 antigen detection method to screen large number of clinical samples for dengue virus isolation. Patient’s serum samples were added onto in vitro tissue culture grown C6/36 or Vero cell lines. Seven days post-infections, culture supernatants were harvested for testing of NS1 antigen by ELISA to confirm the presence of dengue viruses. Significantly higher rate of virus isolation was observed in Vero cells at 7 days post-infection in comparison to C6/36 cells. NS1 antigen could be detected earliest at day 3 post-infection in culture supernatants of both C6/36 and Vero cells. Use of Vero cells in a single 35-mm culture dishes resulted in 69.7% DENV isolations from NS1 alone positive samples. The method is economical, easy to perform, and useful in handling large number of clinical samples and will be of value in virus surveillance, especially post-vaccination. Keywords Dengue virus . Serotypes . Vero . C6/36 . Isolation . Clinical samples
Introduction Dengue is a mosquito-borne viral disease affecting more than 390 million individuals every year in more than 125 countries across the world [1, 2]. In recent years, dengue infection has spread to non-endemic countries, such as Afghanistan, European Union, Chile, and Japan, where several cases of autochthonous dengue outbreak have been reported [3]. India contributes to 34% of dengue infection, and still, the number of officially reported dengue cases is grossly underreported [1, 2]. Efforts are therefore needed for accurate diagnosis of dengue infection. The presence of four distinct serotypes and variable circulation of individual serotypes in different areas and over time coupled with continuous evolution of DENV strains necessitate active dengue virus surveillance. This article is part of the Topical Collection on Medicine * Vidya A. Arankalle [email protected] 1
Department of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Katraj, Pune, Maharashtra 411043, India
Such surveillance is of special significance for dengue infection wherein disease severity is enhanced during secondary dengue infection with a different serotype. At present, a range of laboratory diagnostic methods are available for dengue diagnosis. Depending on the time lapse between appearance of first clinical symptoms and
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