Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
Abstract
:1. Introduction
2. Experimental Section
2.1. Bat Specimens
Species | Abbreviation | Scientific Name | Numbers Tested |
---|---|---|---|
Little brown bat | LBB | Myotis lucifugus | 24 |
Northern long-eared bat | NLB | Myotis septentrionalis | 13 |
California bat | CLB | Myotis californicus | 7 |
Long-legged bat | LLB | Myotis volans | 5 |
Western long-eared bat | LEB | Myotis evotis | 12 |
Yuma bat | YUB | Myotis yumanensis | 8 |
Keen’s bat | KEB | Myotis keenii | 14 |
Big brown bat | BBB | Eptesicus fuscus | 25 |
Townsend’s big-eared bat | WEB | Corynorhinus townsendii | 9 |
Hoary bat | HRB | Lasiurus cinereus | 10 |
Eastern red bat | REB | Lasiurus borealis | 9 |
Silver-haired bat | SHB | Lasionycteris noctivagans | 12 |
Tricolored bat * | EPB | Perimyotis subflavus | 3 |
Pallid bat | PAB | Antrozous pallidus | 2 |
Total | 153 |
2.2. Primer and Probe Design
2.3. DNA Extraction
2.4. Slot Blot Analysis
2.5. PCR Amplification for Microarray Analysis
2.6. Microarray
3. Results and Discussion
3.1. Development of a Universal PCR Primer Pair
3.2. PCR for Partial Amplification of the Cytochrome c Oxidase Subunit I (COI) Gene of Canadian Bats
Name | Specific Target | Sequence (5′-3′) | Location 1 |
---|---|---|---|
BB1 2 | BBB | CTGCCCTGAGTCTGCT | 130–145 |
BB2 2 | BBB | GTGGACCTGACCATT | 465–479 |
BB-E 2 | BBB (eastern) | CTTTTATTCGGCGCTTGA | 96–113 |
BB-W 2 | BBB (western) | TTCTGTTCGGCGCCTGA | 97–113 |
EP1 | EPB | CGCACACGCCTTTG | 215–228 |
EP2 2 | EPB | CTTTCTTTTACTCCTAGCAT | 362–381 |
LAS | HRB/REB | TATTATTGGCATCWTC | 370–385 |
HR1 2 | HRB | AGTACCACTGATGATTGG | 284–301 |
HR2 2 | HRB | CCACCTGCCTTGTCC | 555–569 |
RE1 2 | REB | TCTATAACGTTATCGTGAC | 196–214 |
RE2 2 | REB | GGTACCCCTTATRATCG | 284–300 |
SH1 | SHB | GGGCTCTACTTGGAGAT | 172–188 |
SH2 2 | SHB | CAACTGGTTAGTTCCTCTG | 275–293 |
WE1 2 | WEB | TGATTAATCCCACTAATGAT | 279–298 |
WE2 2 | WEB | CTTACATCTGGCTGG | 485–497 |
CL1 2 | CLB | CTGGGAGACGATCAAATT | 180–197 |
CL2 2 | CLB | TTTTATAGTTATGCCAATCAT | 239–259 |
LB1 2 | LBB | CGCTGAGCTAGGTCAA | 152–167 |
LB2 2 | LBB | TTTCTATTACTGCTGGC | 363–379 |
LE1 | LEB | TGACATAGCCTTTCC | 308–322 |
LE2 2 | LEB | CTGTCTTACTCCTTCTC | 613–629 |
LL1 | LLB | TGTTGGGGGACGATCAGA | 178–195 |
LL2 2 | LLB | TCTTCTCTCTGCACTTA | 478–494 |
NL1 | NLB | AACTGGGCCAGCCA | 157–170 |
NL2 2 | NLB | ATTCGTTTGGTCCGT | 587–601 |
NL3 2 | NLB | ATTCGTTTGGTCTGT | 587–601 |
KE1 2 | KEB | TCATAGTTATGCCCATTA | 241–258 |
KE2 | KEB | TATGCCCATTATAATTGG | 248–265 |
YU1 | YUB | CCCTTTTAGGGGATG | 175–189 |
YU2 2 | YUB | TATAGTAATGCCGATTATAATC | 242–263 |
PA 2 | PAB | TAATGTAATTGTCACAGCA | 200–218 |
3.3. Evaluation of Probes by Slot Blot Analysis
3.4. Evaluation and Validation of Probes by Microarray
Sample/Probe | Sequence (5′-3′) |
---|---|
REB #116 | TTA TAG TTA TGC CCA TTA 1 |
REB #115 | TTA TAG TCA TGC CCA TTA 1 |
KE1 Probe | TCA TAG TTA TGC CCA TTA 1,2 |
KEB #51 | TCA TAG TTA TGC CCA TTA |
4. Conclusions
Acknowledgments
Conflict of Interest
References
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Lung, O.; Nadin-Davis, S.; Fisher, M.; Erickson, A.; Knowles, M.K.; Furukawa-Stoffer, T.; Ambagala, A. Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada. Microarrays 2013, 2, 153-169. https://doi.org/10.3390/microarrays2020153
Lung O, Nadin-Davis S, Fisher M, Erickson A, Knowles MK, Furukawa-Stoffer T, Ambagala A. Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada. Microarrays. 2013; 2(2):153-169. https://doi.org/10.3390/microarrays2020153
Chicago/Turabian StyleLung, Oliver, Susan Nadin-Davis, Mathew Fisher, Anthony Erickson, M. Kimberly Knowles, Tara Furukawa-Stoffer, and Aruna Ambagala. 2013. "Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada" Microarrays 2, no. 2: 153-169. https://doi.org/10.3390/microarrays2020153