Table 3 Details on RNA extraction, PCR methods and kits and PCR volumes used by laboratories participating in the External Quality Assessment
Laboratory idenification number | Volume used for RNA purification (µl) | Elution volume (µl) | Volume of RNA per RT-PCR (µl) | Kit/apparatus used for extraction | Kit for real-time RT-qPCR | Kit for conventional RT-PCR |
|---|---|---|---|---|---|---|
Reference Laboratory | 200 | 60 | 5 | Qiagen: EZ1 Advanced XL with virus minikit v2.0 (9018708) (automated) | Thermo Fisher Scientific: RT-qPCRone-step SuperScript III Platinum (11732088) | Thermo Fisher Scientific:SuperScript™ III One-Step RT-PCR System with Platinum (10928042) |
1 | 200 | 60 | 5 | Resnova: MAGPURIX VIRAL/Pathogen Nucleic Acids Extraction Kit A (ZP02011) (automated) | Meridian Bioscience: 2 × SensiFAST Probe No-ROX Mix Bioline (BIO-76001) | Meridian Bioscience: MyTaqMix One Step RT-PCR Bioline (MDX062) |
2 | 300 | 50 | 5 | Promega: Maxwell® 16 Viral Total Nucleic Acid Purification Kit (AS1150) (automated) | Promega: GoTaq® Probe 1-Step RT-qPCR System (A6120) | Promega: Access RT-PCR System (A1260) |
3 | 200 | 60 | 5 | Thermo Fisher Scientific: MagMAX™ Viral/Pathogen Ultra Nucleic Acid extraction kit (A42356) (manual) | Thermo Fisher Scientific: RT-qPCRone-step SuperScript III Platinum (11732088) | Thermo Fisher Scientific: SuperScript™ III One-Step RT-PCR System with Platinum (12574018) |
4 | 400 | 100 | 5 | Precision System Science Co.: MagLEAD 12gC (MagDEA Dx SV 400) (automated) | Meridian Bioscience: Inhibitor-Tolerant RT-qPCR Mix (MDX016) | Qiagen: OneStep RT-PCR Kit (210212) |
5 | 140 | 60 | 5 | Qiagen: QiaAmp Viral RNA Mini Lit (52904) (manual) | NZY Tech: NZYSpeedy One-step RT-qPCR Probe Master Mix (MB35201) Thermo Fisher Scientific: Taqman master mix (4444434) | Thermo Fisher Scientific: SuperScript™ IV First-Strand Synthesis System (18091050) Thermo Fisher Scientific: DreamTaq PCR Master Mix (2X) (K1071) |
6 | 200 | 30 | 5 in RT-PCR, 4 in qPCR | Qiagen: RNeasy Mini Kit (74104) (manual) | New England Biolabs: Luna Universal Probe One-Step RT-qPCR Kit (E3006S) | Thermo Fisher Scientific: SuperScript™ III One-Step RT-PCR System with Platinum (10928042) |
7 | 200 | 30 | 5 | Qiagen: RNeasy Mini Kit (74104) (manual) | Thermo Fisher Scientific: RT-qPCRone-step SuperScript III Platinum (11732088) | Thermo Fisher Scientific: SuperScript™ III One-Step RT-PCR (11732020) |
8 | 200 | 60 | 5 | Qiagen: EZ1 Advanced XL with virus minikit v2.0 (9018708) (automated) | Thermo Fisher Scientific: RT-qPCRone-step SuperScript III Platinum (11732088) | Thermo Fisher Scientific: SuperScript™ III One-Step RT-PCR (11732020) |
9 | 200 | 50 | 5 | Qiagen: RNeasy Mini Kit (74104) (manual) | Thermo Fisher Scientific: RT-qPCRone-step SuperScript III Platinum (11732088) | Thermo Fisher Scientific: SuperScript™ III One-Step RT-PCR System with Platinum (12574018) |