Table 1 Oligonucleotide primers used for screening bat samples of Coxiella burnetii, Bartonella spp., Rickettsia spp., Ehrlichia spp. and Anaplasma spp.
From: Coxiella and Bartonella spp. in bats (Chiroptera) captured in the Brazilian Atlantic Forest biome
| Pathogen | Target gene | Oligonucleotide primer | Primer sequence (5′ – 3′) | Amplicon size (bp) | Cycling conditions | Reference |
|---|---|---|---|---|---|---|
| Coxiella burnetii | IS1111 |
Outer primer F Outer primer R Nested primer F Nested primer R |
TATGTATCCACCGTAGCCAGC CCCAACAACACCTCCTTATTC AAGCGTGTGGAGGAGCGAACC CTCGTAATCACCAATCGCTTCGTC |
687 bp 440 bp |
95 °C for 5 min, 40 cycles of 95 °C for 30s, 60 °C for 30s, 72 °C for 1 min, final extension of 72 °C for 7 min 95 °C for 5 min, 30 cycles of 95 °C for 30 s, 66 °C for 30 s, 72 °C for 30 s, final extension of 72 °C for 5 min |
[36] [35, 37] |
| Bartonella spp. | gltA |
Outer primer F Outer primer R |
GCTATGTCTGCVTTCTATCAYGA AGAACAGTAAACATTTCN GTHGG | 731 bp | 95 °C for 10 min, 35 cycles of 95 °C for 30s, 58 °C for 30s, 72 °C for 1 min, final extension of 72 °C for 8 min | [35] |
| Rickettsia spp. | gltA |
Outer primer F Outer primer R Nested primer F Nested primer R |
CATCCTATGGCTATTATGCTTGC TATACTCTCTATG(T/A)AC(A/G)T(A/G)ACC CTTACCGCTATTAGAATGATTGC GAGCGA(T/G)AGCTTCAAG(T/C)TCTAT |
885 bp 572 bp |
95 °C for 10 min, 30 cycles of 95 °C for 30 s, 55 °C for 40 s, 72 °C for 55 s, final extension of 72 °C for 10 min 95 °C for 7 min, 25 cycles of 95 °C for 30 s, 63 °C for 30 s, 72 °C for 35 s, final extension of 72 °C for 10 min |
[35] [35] |
| Ehrlichia spp. /Anaplasma spp. | 16 s rRNA |
Outer primer F Outer primer R |
GGTACCYACAGAAGAAGTCC TGCACTCATCGTTTACAG | 345 bp | 95 °C for 3 min, 35 cycles of 95 °C for 15 s, 55 °C for 30 s, 72 °C for 30s, final extension of 72 °C for 5 min | [38] |