Table 5.: Type of Samples, Sample Volume, Storage Conditions, Pre-Processing, Lysis Conditions and System Used to Capture/Elute RNA, for the Viral Load Assays

Abbott RealTime HIV-1 (m2000rt) COBAS® AmpliPrep/COBAS® TaqMan HIV-1, v2.0 (Roche) NucliSens® EasyQ HIV-1 v1.2 (bioMérieux) VERSANT® HIV-1 RNA 1.0 (kPCR) (Siemens) Artus HIVirus-1 QS-RGQ (Qiagen)
Valid samples Human plasma with ACD solution1 or EDTA, DBS2 Human plasma with EDTA, DBS Human plasma with EDTA, DBS, tissues Human plasma with ACD solution1 or EDTA, DBS Human plasma with EDTA
Sample volume Optimum amount 1.0 ml but uses 0.6 ml. Optimum amount 1.0 ml but uses 0.85 ml. 0.5-1.0 ml 0.7-1.35 ml depending on the type and size of tube 1.2 ml
Pre-processing storage conditions Whole blood: Room Temp. 24 h/2-8°C 24 h
Plasma: Room Temp. 24 h/2-8°C 5 days/-80°C indefinitely
Freeze/thaw once only 		Whole blood: Room Temp. 24 h 6°C
Centrifugation: 20 min
Plasma: Room Temp. 24 h/2-8°C 5 days/-80°C indefinitely
Freeze/thaw up to five times		 Whole blood: Room Temp. 4 h
Plasma: 48 h 2-8°C/-80°C indefinitely
Freeze/thaw up to three times		 Whole blood: Room Temp. 6 h, 2-8°C 24 h
Plasma: 5 days 2-8°C/-80°C indefinitely
Freeze/thaw up to four times		 Whole blood: Room Temp. 6 h
Centrifugation: 20 min
Plasma: 4°C several days/-20°C several weeks/-70°C months and years
Sample preparation prior to lysis Clarify by centrifugation at 2000 g x 5 min, uncap; place in the rack with the code facing outwards Insert the clips with the codes in the rack, add the K-tubes, vortex the samples 3-5 s, pipette the samples into the tubes, place them in the rack Pipette the lysis buffer into the apparatus, pipette the samples into the tubes, mix and add the mixture of silica and internal control Place the samples in the tube rack with the code facing outwards and at the appropriate height Prepare the internal control-RNA carrier mixture, place the samples in the tube rack with the code facing outwards
Lysis Sodium guanidine isothiocyanate Incubate with protease and lysis with guanidine thiocyanate Sodium guanidine isothiocyanate/high-concentration saline Proteinase K and chaotropic buffer Proteinase K
Capture/elution of RNA Magnetic particles/water Capture technique based on generic silica/elution with aqueous solution at high temp. Magnetic silica/water Magnetic silica Magnetic particles

1 ACD- citric acid-citrate-dextrose.
2 DBS: Dried blood spots.