Figure 1

Inoculation loop for sample acquisition and configuration of reaction tubes. (A) A 1 µL inoculation loop is used for oral sample withdrawal and collection. (B) A 0.1 mL PCR tube with a Tween-containing reaction mixture immobilized by agarose is placed inside the cap of the tube and 1 µL of SDS/TE solution is placed at the bottom. A clinical sample withdrawn using the inoculation loop is transferred to the bottom of the tube and denatured by mixing it with the SDS/TE solution using the same loop. Subsequently, the nucleic acid detection mixture Tween 80 is transferred from the cap to the bottom of the tube by centrifugation or shaking. (C) Alternatively, 0.1 mL PCR tube with a dried 1 µL spot of SDS/TE on the sidewall of the tube and 5 µL of a Tween-containing reaction mixture at the bottom can be used (left tube). The inoculation loop is rubbed on the side of the tube to dissolve the SDS/TE mixture (marked with permanent marker) and denature the clinical sample and then used to drive the sample to the bottom of the tube. For systems with optical readout at the bottom of the sample tube (e.g., Biomolecular Systems Mic, Quantabio Q tube or Qiagen RotorGene Q) the SDS/TE solution can be dried on the sidewall and the Tween-containing RT-qPCR reaction mixture can be stored underneath 5 µL of oil (right tube).