"Sample pooling of RNA extracts to speed up SARS-CoV-2 diagnosis using CDC FDA EUA RT-qPCR kit"

Analytical sensitivity and clinical performance of a triplex RT-qPCR assay using CDC N1, N2, and RP targets for SARS-CoV-2 diagnosis

“…All the samples included on the study were tested following an adapted version of the CDC protocol ( Lu et al, 2020 , Interim Guidelines for Collecting, Handling, and Testing Clinical Specimens from Persons for Coronavirus Disease, 2019 ) using AccuPrep Viral RNA extraction kit (Bioneer, South Korea), TaqMan Fast Virus 1-Step Master Mix (Applied Byosistems, USA) and CFX96 thermal cycler (BioRad) ( Freire-Paspuel et al, 2020a , Freire-Paspuel et al, 2020b , Freire-Paspuel et al, 2020c ).…”

High prevalence of SARS-CoV-2 infection among food delivery riders. A case study from Quito, Ecuador

“…Nasopharyngeal swabs were collected on 0.5 mL TE pH 8 buffer for SARS-CoV-2 diagnosis by RT-qPCR following an adapted version of the CDC protocol by using PureLink Viral RNA/DNA Mini Kit (Invitrogen, USA) as an alternate RNA extraction method and CFX96 BioRad instrument ( Interim Guidelines for Collecting, n.d. ; Freire-Paspuel et al, 2020a ; Freire-Paspuel et al, 2020b ; Freire-Paspuel et al, 2020c ; Freire-Paspuel et al, 2020d ; Freire-Paspuel and Garcia-Bereguiain, 2020 ). Briefly, the CDC designed RT-qPCR FDA EUA 2019-nCoV CDC kit (IDT, USA) is based on N1 and N2 probes to detect SARS-CoV-2 and RNase P as an RNA extraction quality control ( Interim Guidelines for Collecting, n.d. ; Freire-Paspuel et al, 2020a ; Freire-Paspuel et al, 2020b ; Freire-Paspuel et al, 2020c ; Freire-Paspuel et al, 2020d ; Freire-Paspuel and Garcia-Bereguiain, 2020 ). Also, negative controls (TE pH 8 buffer) were included as control for carryover contamination, one for each set of RNA extractions, to guarantee that only true positives were reported.…”

Low clinical performance of the Isopollo COVID-19 detection kit (M Monitor, South Korea) for RT-LAMP SARS-CoV-2 diagnosis: A call for action against low quality products for developing countries

“…The viral loads detailed in Supplementary Material Tables S1 and S 2 were calculated running a calibration curve with the 2019-nCoV N positive control (IDT, USA). The LoD for the CDC protocol was set at 1000 viral RNA copies per milliliter of sample (or 5 RNA copies/μl of RNA extraction solution) in previous studies ( Lu et al, 2020 , Freire-Paspuel et al, 2020a , Freire-Paspuel et al, 2020b , Freire-Paspuel et al, 2020c , Freire-Paspuel and Garcia-Bereguiain, 2020a , Freire-Paspuel et al, 2020 , Freire-Paspuel and Garcia-Bereguiain, 2020b ). Although the LoD could not be calculated for the Isopollo COVID-19 detection kit, as described in the Methods section, even for samples with viral loads above 100 000 RNA copies/ml (500 RNA copies/μl of RNA extraction solution), only 81 out of 88 (92.04%) samples were also positive with the Isopollo COVID-19 detection kit.…”

“…All of the samples included in the study were tested following an adapted version of the CDC protocol reported previously by our laboratory ( Freire-Paspuel et al, 2020b , Freire-Paspuel et al, 2020c , Freire-Paspuel and Garcia-Bereguiain, 2020a , Freire-Paspuel et al, 2020 , Freire-Paspuel and Garcia-Bereguiain, 2020b , Freire-Paspuel et al, 2020d ).…”