Volume 7, Number 6—December 2001
Rapid Identification of Bordetella pertussis Pertactin Gene Variants Using LightCycler Real-Time Polymerase Chain Reaction Combined with Melting Curve Analysis and Gel Electrophoresis
|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.