TABLE 2.
HEV and internal control RT-qPCR primers and Pleiades probes
| Target and primer or probe | Nucleotide sequencea | Concn (1×) (μM) |
|---|---|---|
| HEV ORF 2/3 | ||
| HEV forward primer | 5′-AATAAATCATAAGGTGGTTTCTGGGGTGAC-3′ | 0.250 |
| HEV reverse primer | 5′-AATAAATCATAAGGGGTTGGTTGGATGAA-3′ | 1.000 |
| HEV probe | 5′-MGB-FAM-G*TGATTCTCAGCCCTTCG-NFQ-3′ | 0.400 |
| MS2 RNA internal control | ||
| MS2 forward primer | 5′-CCA*TCAAA*GTCGA*GGTGCCTAAAGTG-3′ | 0.075 |
| MS2 reverse primer | 5′-ACGAACGCCATGCGGCTACAGGAAGCTC-3′ | 0.075 |
| MS2 probe | 5′-MGB-AP525-G*CTGTTGGTGGTGTAGAGC-NFQ-3′ | 0.200 |
a
Underlined nucleotides represent 5′ flaps added to enhance RT-qPCR amplification (31). G*and A* represent super G and super A bases (ELITechGroup Inc.) incorporated to enhance stability (32). AP525, proprietary fluorescent dye (ELITechGroup Inc.); FAM, 6-carboxyfluorescein; MGB, minor grove binder; NFQ, nonfluorescent quencher; ORF, open reading frame.