Nucleic acid based diagnostic techniques are rou-tinely used for the detection of infectious agents. Most of theseassays rely on nucleic acid extraction platforms for the extrac-tion and purification of nucleic acids and a separate real-timePCR platform for quantitative nucleic acid amplification tests(NATs). Several microfluidic lab on chip (LOC) technologieshave been developed, where mechanical and chemical methodsare used for the extraction and purification of nucleic acids.Microfluidic technologies have also been effectively utilizedfor chip based real-time PCR assays. However, there are fewexamples of microfluidic systems which have successfully in-tegrated these two key processes. In this study, we have imple-mented an electro-actuation based LOC micro-device that le-verages multi-frequency actuation of samples and reagentsdroplets for chip based nucleic acid extraction and real-time,reverse transcription (RT) PCR (qRT-PCR) amplification fromclinical samples. Our prototypemicro-device combines chem-ical lysis with electric field assisted isolation of nucleic acid in afour channel parallel processing scheme. Furthermore, a fourchannel parallel qRT-PCR amplification and detection assay isintegrated to deliver the sample-to-detection NAT chip. TheNAT chip combines dielectrophoresis and electrostatic/electrowetting actuation methodswith resistive micro-heatersand temperature sensors to perform chip based integratedNATs. The two chip modules have been validated using differ-ent panels of clinical samples and their performance comparedwith standard platforms. This study has established that ourintegrated NAT chip system has a sensitivity and specificitycomparable to that of the standard platforms while providingup to 10 fold reduction in sample/reagent volumes

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