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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

Johanna Mäkinen*†Comments to Author , Matti K. Viljanen*, Jussi Mertsola†, Heikki Arvilommi*, and Qiushui He*
Author affiliations: *National Public Health Institute, Department in Turku, Finland;; †Turku University Central Hospital, Turku, Finland

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Table 1

Primers and probes used in study of Bordetella pertussis pertactin gene variants

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.

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