TABLE 2.
Oligonucleotides used for home-brew molecular assay
| Assay step and primer (sequence) or product | Virus and positions amplifieda | Length (nucleotides) | G+C content (%) | Melting temp (°C) |
|---|---|---|---|---|
| Initial PCR | ||||
| Primers | ||||
| Forward (5′-TGCTCCTACAACAAGTC) | HSV-1, 138845–138861; HSV-2, 141422–141438 | 17 | 47.1 | 43.8 |
| Reverse (5′-CGGTGCTCCAGGATAAA) | HSV-1, 139028–139044; HSV-2, 141625–141641 | 17 | 52.9 | 51.4 |
| Product | HSV-1, 138845–139044; HSV-2, 141422–141641 | 200 | ||
| Nested PCR | ||||
| Primers | ||||
| Forward (5′-ATCCGAACGCAGCCCCGCTG) | HSV-1, 138878–138897; HSV-2, 141475–141494 | 20 | 70.0 | 70.1 |
| Reverse (5′-TCTCCGTCCAGTCGTTTATCTTC) | HSV-1, 138997–139019; HSV-2, 141594–141616 | 23 | 47.8 | 57.3 |
| Product | HSV-1, 138878–139019; HSV-2, 141475–141616 | 142 |