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Table 3 Primer and probe sequences for amplification of C. pneumoniae, M. pneumoniae, macrolide resistance associated mutations in M. pneumoniae and human beta-globin DNA, as well as the primer sequences used for sequencing of M. pneumoniae 23S rRNA

From: Development of a multiplex real-time PCR assay for detection of Mycoplasma pneumoniae, Chlamydia pneumoniae and mutations associated with macrolide resistance in Mycoplasma pneumoniae from respiratory clinical specimens

Primer/probe

Sequence 5′->3′

Gene target (reference)

MP Fw

ATGTACTATCAGCAAAAGCTCAGTATGG

M. pneumoniae Hyppthetical protein C985_0367

(GeneBank accesion no: AGC04275.1)

(this publication)

MP Rev

CCACATACCGCTTTAAGTTAGCAA

MP probea

Cy5-CTAACCAAAACAGCCCTTCAACGGCA-Iowa black RQ-Sp

CP Fw

AAGGGCTATAAAGGCGTTGCT

C. pneumoniae ompA

(Mannonen et al. 2004)

CP Rev

TGGTCGCAGACTTTGTTCCA

CP probea

Tx Red-TCCCCTTGCCAACAGACGCTGG-Iowa black RQ-Sp

BG Fw

GGTTGGGATAAGGCTGGATTATT

Homo sapiens beta-globin chain

(GeneBank accesion no: AY260740.1)

(this publication)

BG Rev

CAGGAGCTGTGGGAGGAAGA

BG probea

JOE/ZEN-CAAGCTAGGCCCTTTTGCTAATCATGTTCA-Iowa black FQ

MR–MP Fw

GCTATAGACTCGGTGAAATCCAGG

M. pneumoniae gene for 23S rRNA (GeneBank accesion no: gi|288509|emb|X68422.1|)

(this publication)

MR–MP Rev

GCTACAGTAAAGCTTCACTGGGTC

MR–MP SimpleProbe®

GCGCA XI ACGGGACGGAAAGAC

CARDS TX Fw

TTTGGTAGCTGGTTACGGGAAT

CARDS toxin gene

(Winchell et al. 2008)

CARDS TX Rev

GGTCGGCACGAATTTCATATAAG

CARDS TX probea

TGTACCAGAGCACCCCAGAAGGGCT

M. pneumoniae Fw diagnostic PCR

AAGGACCTGCAAGGGTTCGT

16SrRNA

(van Kuppeveld et al. 1992)

M. pneumoniae Rev diagnostic PCR

Biotin-CTCTAGCCATTACCTGCTAA

M. pneumoniae probe diagnostic PCR

ACTCCTACGGGAGGCAGCAGTA-digoxigenin

MR Seq Fwb

AACTATAACGGTCCTAAGGTAGCG

23srRNA

(Wolff et al. 2008)

MR Seq Revb

GCTCCTACCTATTCTCTACATGAT

  1. aDually labelled hydrolysis probe
  2. bSequencing primer