Table 5.
Sequences of primers and probes used in conventional PCR and RQ-PCR assays
| Oligonucleotides (5′ → 3′)a | Reference positionsb | Temperature | Size (bp) PCR product | |
|---|---|---|---|---|
| PCR, SV40 early region | ||||
| SVEARL1: | CAGCAGTAGCCTCATCATCAC | nt 4463–4483 | 52 | 669, viral DNA |
| SVEARL2: | GCAGCTAATGGACCTTCTAGGTCT | nt 5131–5108 | 602, small t-ag cDNA 322, large T-ag cDNA |
|
| PCR, SV40 late region | ||||
| SVLATE1: | CTCCGTTAAGGTTCGTAGGTC | nt 364–384 | 52 | 1283, viral DNA |
| SVLATE2: | CCTCAGTGAAGCTGTCTACTC | nt 1646–1626 | 346, 16S RNA cDNA | |
| RQ-PCR, SV40 early region, SVPENT primers and probe | ||||
| FP: | GATGGCATTTCTTCTGAGCAAA | nt 4486–4507 | N/A | N/A |
| RP: | GAATGGGAGCAGTGGTGGAA | nt 4550–4531 | ||
| Probe: | FAM-CAGGTTTTCCTCATTAAA-MGB | nt 4509–4526 | ||
| RQ-PCR, SV40 late region, SVVP1 primers and probe | ||||
| FP: | GGGCCCTTGTGCAAAGC | nt 2258–2274 | N/A | N/A |
| RP: | GTTGGTAAACAGCCCACAAATG | nt 2323–2302 | ||
| Probe: | VIC-ACAGCTTGTATGTTTCTG-MGB | nt 2277–2294 | ||
| RQ-PCR, human 18S ribosomal RNA | ||||
| FP: | TCGAGGCCCTGTAATTGGAA | N/A | N/A | N/A |
| RP: | CCCTCCAATGGATCCTCGTT | |||
| Probe: | FAM-AGTCCACTTTAAATCCTT-MGB | |||
a
FP = forward primer; RP = reverse primer.
b
Reference nucleotide positions in SV40 strain 776 (SV40-776).
c
PCR annealing temperature.
N/A = not applicable.