Here, we developed a one-step detection method for HIV virus based on Flinders Technology Associates®(FTA®) card and isothermal nucleic acid amplification. This method aims to establish a simple, rapid and sensitive disease detection system by sample-to-answer nucleic acid pretreatment, amplification, and detection. Blood sample could be rapid lysed and nucleic acid such as DNA and RNA would be absorbed and enriched to the FTA® card. Moreover, this method could efficiently protect RNA from degradation because the special design of FTA® card which has high anti-interference capacity to microorganism, UV and nuclease interference. The enriched nucleic acid was directly used for the subsequent one-step strand exchange isothermal nucleic acid amplification (SEA) by adding the round slices of the FTA® card to the reaction system. SEA could achieve one-step detection of RNA, depending on single-stranded denatured bubbles and innate reverse transcriptase activity of Bst DNA polymerase. Compared to traditional polymerase chain reaction (PCR), no extra reverse transcription step was need, which greatly reduce the detection time. The method could realize pre-treatment of sample and enrichment of nucleic acid in 30 minutes and SEA could be finished within 30 minutes. The detection method is simple, fast and needs no large instruments, making it well suitable for high throughout and high efficiency screening of blood samples.

One-step assay for detection of HIV virus with sample-in-answer-out capability

255th National Meeting (2018)

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