Table 1.
Primers used for multiplex RT-PCR and hemi-nested PCR
| Virus | Primers | Sequence (5′ → 3′) | Gene | Amplicon size (bp) | Melting temp (°C) | Authors | |
|---|---|---|---|---|---|---|---|
| Primers multiplex 2 | |||||||
| RT-PCR | hRSV | vrs P1 | GGA ACA AGT TGT TGA GGT TTA TGA ATA TGC | Nucleocapsid | 279 pb | 60 | Cane and Pringle (1991) |
| vrs P2 | TTC TGC TGT CAA GTC TAG TAC ACT GTA GT | 55 | |||||
| Influenza A virus | mia 1 | CAG AGA CTT GAA GAT GTC TTT GCT GG | Matrix protein | 212 | 68 | Donofrio et al. (1992) | |
| mia 2 | GCT CTG TCC ATG TTA TTT G | 68 | |||||
| Influenza B virus | mib 1 | AAA ATT ACA CTG TTG GTT CGG TG | Matrix protein | 362 | 70 | Donofrio et al. (1992) | |
| mib 2 | AGC GTT CCT AGT TTT ACT TG | 72 | |||||
| hMPV | hmpv 1 | CCC TTT GTT TCA GGC CAA | Matrix protein | 416 | 54 | This study | |
| hmpv 2 | GCA GCT TCA ACA GTA GCT G | 58 | |||||
| Hemi-nested | hRSV | vrs i | GGT GTA CCT CTG TAC TCT C | Nucleocapsid | 180 | 58 | This study |
| Influenza A virus | mia 3 | CTC TGA CTA AGG GGA TTT TG | Matrix protein | 130 | 58 | This study | |
| Influenza B virus | mib 3 | CAT GAA ARC TCA CAC ATC T | Matrix protein | 260 | 53 | This study | |
| hMPV | hmpv 3 | AGG CCA ACA CAC CAC CAG | Matrix protein | 410 | 58 | This study | |
| Primers multiplex 2 | |||||||
| RT-PCR | Parainfluenza virus 1 | PIS1+ | CCG GTA ATT TCT CAT ACC TAT G | Hemagglutinin- | 317 pb | 48 | Echevarria et al. (1998) |
| PIS1− | CCT TGG AGC GGA GTT GTT AAG | Neuraminidase | 51 | ||||
| Parainfluenza virus 2 | PIP2+ | AAC AAT CTG CTG CAGCAT TT | Hemagglutinin- | 507 | 56 | Echevarria et al. (1998) | |
| PIP2− | ATG TCA GAC AAT GGG CAA AT | Neuraminidase | 56 | ||||
| Parainfluenza virus 3 | Para3.1 | CTC GAG GTT GTC AGG ATA TAG | Hemagglutinin- | 189 | 46 | Karron et al. (1994) | |
| Para3.2 | CTT TGG GAG TTG AAC ACA GTT | Neuraminidase | 48 | ||||
| Parainfluenza virus 4 | PIP4+ | CTG AAC GGT TGC ATT CAG GT | Phosphoprotein | 451 | 60 | Aguilar et al. (2000) | |
| PIP4− | TTG CAT CAA GAA TGA GTC CT | 56 | |||||
| Internal control | GAPDH1 | TCA TCC ATG ACA ACT TTG GTA TCG TG | GAPDH | 564 | 59 | Gueudin et al. (2003) | |
| GAPDH2 | CTC TTC CTC TTG TGC TCT TG | 60 | |||||
| Hemi-nested | Parainfluenza virus 1 | PiS1i | AGC TGC AGG AAC AAG GGG | Hemagglutinin-N | 261 | 58 | This study |
| Parainfluenza virus 2 | Para2i | CTA GCT GAA CTG AGA CTT G | Hemagglutinin-N | 340 | 56 | This study | |
| Parainfluenza virus 3 | Para3i | GCT AGA GAA CAT GAC TTC C | Hemagglutinin-N | 145 | 56 | This study | |
| Parainfluenza virus 4 | Pi4i | GTC TGA TCC CAT AAG CAG C | Phosphoprotein | 390 | 58 | This study | |
| Primers multiplex 3 | |||||||
| RT-PCR | hRV | SRHI1 | GCA TCI GGY ARY TTC CAC CAC CAN CC | VP4/VP2/5’NC | 549 | 62 | Savolainen et al. (2002) |
| SRHI2 | GGG ACC AAC TAC TTT GGG TGT CCG TGT | 65 | |||||
| HCoV 229E | MD1 | TGG CCC CAT TAA AAA TGT GT | Gene M | 573 | 60 | Vabret et al. (2001) | |
| MD3 | CCT GAA CAC CTG AAG CCA AT | 60 | |||||
| HCoV OC43 | MF1 | GGC TTA TGT GGC CCC TTA CT | Gene M | 335 | 58 | Vabret et al. (2001) | |
| MF3 | GGC AAA TCT GCC CAA GAA TA | 58 | |||||
| Influenza C virus | CHAA | ACA CTT CCA ACC CAA TTT GG | Hemagglutinin-esterase | 485 | 58 | Zhang and Evans (1991) | |
| CHAD | CCT GAC AGC AAC TCC CTC AT | 62 | |||||
| Hemi-nested | hRV | Nestrhi1 | ATG GGN GCW CAN GTN TCH ANH CA | VP4/VP2/5′NC | 450 | 53 | This study |
| HCoV 229E | MD2i | CCG TAT CAA CAC TCG TTA TGT GG | Gene M | 230 | 54 | This study | |
| HCoV OC43 | MF2i | CTC CAA AAA CTT CCA GTT C | Gene M | 170 | 54 | This study | |
| Influenza C virus | MICi | GAG GAT GTG GCA ACT ACT | Haemagglutinin-E | 391 | 54 | This study | |