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Development of Low Cost Two-Step Reverse Transcription-Quantitative Polymerase Chain Reaction Assays for Rotavirus Detection in Foul Surface Water Drains.

Food Environ Virol.. 2013-06;  5(2):126-133
J. W. Chung, J. W. Foppen, P. N. L. Lens. Department of Environmental Engineering and Water Technology, UNESCO-IHE Institute for Water Education, P.O. Box 3015, 2601, DA, Delft, The Netherlands
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摘要

Commercial kits to determine RNA concentrations are expensive, and sometimes too expensive for laboratories working with tight budgets, especially those in developing countries. We developed, tested, and evaluated two home-made two-step reverse transcription-quantitative polymerase chain reaction assays aimed to detect rotavirus in surface water samples. A commercial one-step master kit was used for comparison. Our results indicated that the efficiency of the home-made assays was comparable to the commercial kit. Furthermore, the lowest detection limit of all assays corresponded to 10-0.2 TCID50 (50 % tissue Culture Infective Dose) per ml. The home-made assays were able to detect rotavirus concentrations in com... More

关键词

Rotavirus; RT-qPCR; Surface water; Slums; Kampala; Uganda
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