Volume 7, Number 6—December 2001
Research
Rapid Identification of Bordetella pertussis Pertactin Gene Variants Using LightCycler Real-Time Polymerase Chain Reaction Combined with Melting Curve Analysis and Gel Electrophoresis
Table 1
Primer/probe | Sequence (5'-3')a | Positionb |
---|---|---|
QJF3c | GCT GGT GCA GAC GCC AGT | 1578-1595 |
QJR1c | CCG ATA TCG ACC TTG CC | 1649-1633 |
QH8Fd | CTG CAG CGC GCG ACG ATA | 757-774 |
QH2Rd | ATT GCC GTG CGG TGC GGA CAA | 1026-1006 |
QJ1e | CCG GCG GTG CGG TTC C-F | 809-824 |
QJ2e | LC Red 640-CGG TGG TGC GGT TCC C-P | 825-840 |
aModifications of primer or probe are boldfaced or underlined.
bPosition numbers indicate the position of bases relative to the first start codon of prn1.
cPrimers used in the real-time allele-specific amplification. QJF3 contained a specific mismatch G (underlined) at the 3'end that does not complement the published sequences of any prn type. The T (boldfaced) at the 3'end (corresponding to the nucleotide 1595) is complementary to prn1-5. Primer QJF3 has two mismatches with prn6-8.
dPrimers used in fluorescence resonance energy transfer (FRET) probe assay.
eProbes used in FRET probe assay. Boldfaced T is complementary to C to T transition specific for prn1. QJ1 was labeled with fluorescein at the 3' end and QJ2 with LC Red at 5'end and phosphorylated at the 3'end.