title: Development of low cost two-step reverse transcription-quantitative polymerase chain reaction assays for rotavirus detection in foul surface water drains.
abstract: 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 complex surface waters in a slum area in Kampala (Uganda) and their performance was comparable to the commercial kit. The total costs of the two home-made assays was 11 times less than the selected commercial kit. Although preparing home-made assays is more time consuming, the assays can be useful for cases in which consumable costs are more important than personnel costs.
authors.affiliation: Department of Environmental Engineering and Water Technology, UNESCO-IHE Institute for Water Education, P.O. Box 3015, 2601 DA, Delft, The Netherlands. w.chung@unesco-ihe.org
authors.forename: J W
authors.initials: JW
authors.lastname: Chung
authors.forename: J W
authors.initials: JW
authors.lastname: Foppen
authors.forename: P N L
authors.initials: PN
authors.lastname: Lens
mesh_terms: D003365:Costs and Cost Analysis; D003906:Developing Countries / Q000191:economics; D012367:RNA, Viral / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic / Q000191:economics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D014454:Uganda; D014867:Water / Q000032:analysis; D014871:Water Microbiology
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D014867:Water