TABLE 1.
qPCR assay primers, probes, DNA targets, and assay characteristicsa
| DNA target | Primer or probe (sequence, 5′→3′) | Product (bp) | Reference | ||
|---|---|---|---|---|---|
| LODb | LOQc | ||||
| IS900 | IS900F (CCGCTAATTGAGAGATGCGATTGG) | 230 | 1.8 | 1.8 | 13 |
| IS900R (ATTCAACTCCAGCAGCGCGGCCTC) | |||||
| IS900P (6-FAM-TCCACGCCCGCCCAGACAGG-TAMRA) | |||||
| Target 251 | 251F (GCAAGACGTTCATGGGAACT) | 200 | ND | ND | 21 |
| 251R (GCGTAACTCAGCGAACAACA) | |||||
| 251P (6-FAM-CTGACTTCACGATGCGGTTCTTC-TAMRA) | |||||
a
FAM, 6-carboxyfluorescein; TAMRA, 6-carboxytetramethylrhodamine; ND, not determined.
b
The limit of detection (LOD) of the IS900 qPCR assay was defined as the lowest copy number resulting in a CT of <40, determined from six independent dilution series.
c
The limit of quantification (LOQ) was defined as the lowest copy number per assay yielding a coefficient of variation (CV) of less than 25% (33).