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Volume 9, Number 12—December 2003

Research

Human Monocytotropic Ehrlichiosis, Missouri

Juan P. Olano*Comments to Author , Edwin Masters†, Wayne Hogrefe‡, and David H. Walker*
Author affiliations: *University of Texas Medical Branch, Galveston, Texas, USA; †Premier Family Physicians, Cape Girardeau, Missouri, USA; ‡Focus Technologies, Cypress, California, USA

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Table 1

List of PCR primers used in this study for amplification of ehrlichial DNA sequences from blood specimens. Cape Girardeau, Missouri, 1997–1999

Target gene Outside primer pair Nested primer pair Cycles: T° (time)* for outside primers Cycles: T° (time)* for nested primers
16S rRNA subunit gene. E. chaffeensis
ECB
5′CGTATTACCGCGGCTGCTGGA-3′
ECC
5′AGAACGAACGCTGGCGGCAGCC-3′
HE1 5′’CAATTGCTTATAACCTTTTCCTTATAAAT-3′
HE3
5′TATAGGTACCGTCATTATCTTCCCTAT-3'
94 (60) 45 (120) 72(60)
94 (60) 55 (120) 72(60)
120-kDa protein gene
E. chaffeensis
PXCF3
5′GAGAATTGATTGTGGAGTTGG-3′
PXAR4
5′ACATAACATTCCACTTTCAAA-3′
PXCF3b
5′-CAGCAAGAGCAAGAAGATGAC-3′
PXAR5
5′ATCT′
94(60) 48(120) 72(60)
94(60) 54(120) 72(60)
nadA gene
E. chaffeensis
ECHNADA1
5′-TCATTTCGTGCTTTCTTATTG-3′
PXCR6
5′-CAAACGCATATG TGGGCA-3′
NADPCR
5′ACGTCATTTGGCTCAGGA-3′
PXCR7
5′-TGTCGATCCAATGAAAT GAGC-3′
94(60) 48(120) 72(60)
94(60) 48(120) 72(60)
16S rRNA subunit gene. A. phagocytophilum PC5
5′-TACCTTGTTACGACTT-3′
Pomod
5′-AGAGTTTGATCCTGG-3′ GE9f
5′-AACGGATTATTCTTTATAGCTTGCT-3′
GE10r
5′-GGAGATTAGATCCTCTTAACGGAA-3′ 94(60) 38(120) 72(60) 94(60) 60(120) 72(60)

a Temperature sequence: Denaturing, annealing and synthesis. Time given in seconds. All polymerase chain reactions (PCR) were performed for 35 cycles.

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