Evaluating the Impact of Presence of Organic Matter on Environmental Samples and Sample Processing Technique on RNA Detection of PEDV

Environmental sampling has become a commonly accepted diagnostic technique for means of identifying breaks in biosecurity. However, environmental samples have yet to be validated reverse transcriptase real-time PCR (qRT-PCR) analysis and there is no standardization sample processing. Therefore, the objective this project was evaluate different types samples, whether processing prior qRT-PCR would impact results. Steel coupons were inoculated with PEDV conditions, then environmentally swabbed using cotton gauze. Treatments arranged as 5 × 4 factorial five treatments contamination four Samples processed ways: pre-qRT-PCR processing, centrifuging, syringe filtering, centrifuging filtering determine if pre-sample impacted cycle threshold (Ct) value. Once processed, they submitted analysis. Results reported proportion positive resulting Ct If had detectable RNA, assigned value 45. For values, an surface (P < 0.0001) interaction indicating that type way sample. pure virus PBS, difference values between techniques (PP 0.05). fecal contamination, not or only greater amounts RNA detected compared filtered centrifuged organic matter all other Main effects also significant. inoculation type, PBS feces inoculation, while dirt intermediate. greatest amount unprocessed being In summary, are particularly dirty, will