TABLE 2.
Characteristics of the HDV RNA species-specific RT-PCR procedures
| Protocol | RNA detected | RNAs transfected | RT primera | Reverse transcriptase enzyme | Temp of RT reaction (°C) | PCR primera (F/R)b | Sensitivity | Specificityc (discrimination) |
|---|---|---|---|---|---|---|---|---|
| mI-S | mRNA-I | G-RNA and mRNA-I/II | Oligo(dT)20 | SuperScript III | 55 | Tagm-T20-F/mI-R | 103 | 6 log10 |
| mI/II-S | mRNA-I/II | G-RNA and mRNA-I/II | Oligo(dT)20 | SuperScript III | 55 | mII-R/mI-F | —d | — |
| G-S | G-RNA | AG-RNA and mRNA-I/II | Tagged GS | rTth | 70 | TagG-F/GS-R | 103 | 5 log10 |
| AG-S | AG-RNA | G-RNA and mRNA-I/II | Tagged AGS | rTth | 70 | TagAG-F/AGS-R | 103 | 5 log10 |
The sequences of the RT-PCR primers are listed in Table 1.
F, forward primer; R, reverse primer.
The specificity for the mI-S protocol is defined as the log difference between the titer at which the mRNA-I was detectable and the titer at which the G-RNA or AG-RNA generated the first positive signal. The specificity for the G-S protocol is defined as the log difference between the titer at which the G-RNA was detectable and the titer at which the AG-RNA generated the first positive signal. The specificity for AG-S protocol is defined as the log difference between the titer at which the AG-RNA was detectable and the titer at which the G-RNA generated the first positive signal.
—, not determined.