Abstract

MORIER DIAZ, Luis et al. Obtainment of CLA-HT cell subline and study of its susceptibility for the isolation and multiplication of the dengue virus. Rev Cubana Med Trop [online]. 2014, vol.66, n.3, pp.424-432. ISSN 0375-0760.

Introduction: the identification of viral serotypes is an important issue for dengue diagnosis. Despite the existence of other identification methods, the viral isolation in cell cultures and determination by immunofluorescent technique remain as the most used. Therefore the search for more sensitive cellular systems has been a repeated topic during many years for dengue virus and other agents. Objective: to obtain a cell subline from CLA-1 clone (Aedes pseudoscutellaris), that grows at 28 °C, capable of multiplying at 33 °C (CLA-HT)and to evaluate its usefulness for Dengue virus isolation and identification. Methods: by using a CLA-1 clone of the AP-61(Aedes pseudoscutellaris) cell line, it was possible to obtain a strain or cell subline CLA-HT capable of growing at 33 oC (CLA-HT) through a temperature selection method. Its sensitivity for the multiplication of the 4 dengue virus serotypes and its efficiency for direct virus isolation from acutely ill patients' sera were studied in comparison to C6/36 HT cell line (A. albopictus). Results: four dengue virus serotypes grew in CLA-HT cell subline but some serotypes were detected later in CLA-HT than in C6/36 HT. For dengue isolation from serum samples taken in acutely ill patients, the CLA-HT subline detected 40 % positive samples whereas C6/36 HT did 50 %. Discussion: CLA-HT is able to detect all dengue virus serotypes from 96 hours on post inoculation. It makes the new cell line useful for research as an alternative system and the direct isolation efficiency is good to be applied in large outbreaks. The most important advantage of CLA-HT is the possibility of giving rapid answer in emergency situations in low resource laboratories since it does not require special incubation conditions with CO2. Conclusions: a CLA-HT subline was obtained from CLA-1 line and it grows at 33 oC and capable of detecting 4 dengue virus serotypes. Its isolation efficiency is slightly lower than that of C6/36 HT subline, but it may be used as an alternative system for dengue virus isolation, mainly in resource-poor laboratories that do not have the optimal conditions for incubation.

Keywords : dengue virus; cell subline; viral detection.

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