Volume 9, Number 12—December 2003
Research
Human Monocytotropic Ehrlichiosis, Missouri
Juan P. Olano*
, Edwin Masters†, Wayne Hogrefe‡, and David H. Walker*
, Edwin Masters†, Wayne Hogrefe‡, and David H. Walker*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.
