Fig. 5: High-sensitivity PCR-free detection of SARS-CoV-2 RNA extracted from BSNFs-chip. | Nature Communications

Fig. 5: High-sensitivity PCR-free detection of SARS-CoV-2 RNA extracted from BSNFs-chip.

From: Biporous silica nanostructure-induced nanovortex in microfluidics for nucleic acid enrichment, isolation, and PCR-free detection

Fig. 5: High-sensitivity PCR-free detection of SARS-CoV-2 RNA extracted from BSNFs-chip.

a Workflow of the luminescence resonance energy transfer (LRET) assay for SARS-CoV-2 RNA collected using BSNFs-chip. b Schematic diagram of the full-length SARS-CoV-2 genome map showing the target gene site for the LRET assay. c Analytical sensitivity of the LRET assay for serial diluted SARS-CoV-2 RNA. a.u., arbitrary units. Data are expressed as mean ± SD (n = 3 independent experiments). p values: p = 0.019 (10−1 PFU), p = 1.37E-7 (100 PFU), p = 3.49E−12 (101 PFU), p = 5.02E-15 (102 PFU), p = 1.42E-18 (103 PFU), p = 8.18E-19 (104 PFU). d–g Diagnostic accuracy of the LRET-based detection towards clinical samples, including 20 SARS-CoV-2 positive samples and 10 negative samples. (d, e) The LRET assay integrated with the BSNFs-chip shows significantly higher sensitivity (p = 0.000019) than with a conventional extraction method (p = 0.009). Data are expressed as mean (n = 3 independent experiments). f, g The LRET assay integrated with the BSNFs-chip has significantly higher accuracy (sensitivity of 100% and area under the ROC curve (AUC) of 1) than that integrated with the conventional method (sensitivity of 30% and AUC of 0.597). p values in (c–e) were determined by a two-tailed unpaired t-test; *p  <  0.05, **p  <  0.01, ***p  <  0.001 and ****p  <  0.0001. The cut-off value (0.0395) was determined by applying optimal combinations of clinical sensitivity and specificity from receiver operator characteristic (ROC) curve based on the Youden index point. Source data are provided as a Source data file.

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