A real-time combined polymerase chain reaction assay for the rapid detection and differentiation of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii
- PMID: 16263231
- DOI: 10.1016/j.diagmicrobio.2005.06.019
A real-time combined polymerase chain reaction assay for the rapid detection and differentiation of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii
Abstract
A rapid real-time polymerase chain reaction (PCR) assay capable of the simultaneous detection and differentiation of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii was developed using the LightCyclertrade mark instrument (Roche Applied Sciences, Indianapolis, IN). The assay targets the operon groEL of the heat shock protein. Base pair mismatches in amplified DNA in regions of detection probe hybridization allowed organism differentiation by melting curve analysis. The analytical sensitivity was at least 10 copies per reaction. DNA extracts from 59 specimens previously confirmed positive for A. phagocytophilum (n = 37), E. chaffeensis (n = 19), or E. ewingii (n = 3) were used to evaluate the assay. All of the specimens positive for 1 of the 3 organisms by conventional PCR were likewise positive by the LightCycler method. Sensitivity and specificity were at least 100% compared with conventional PCR. This assay provides a rapid method for the detection and differentiation of the causative agents of human ehrlichiosis in the United States.
Similar articles
-
Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay.J Clin Microbiol. 2001 Nov;39(11):3871-6. doi: 10.1128/JCM.39.11.3871-3876.2001. J Clin Microbiol. 2001. PMID: 11682500 Free PMC article.
-
Polymerase chain reaction evidence of Ehrlichia chaffeensis, an etiologic agent of human ehrlichiosis, in dogs from southeast Virginia.Am J Vet Res. 1996 Aug;57(8):1175-9. Am J Vet Res. 1996. PMID: 8836370
-
Detection of Ehrlichia chaffeensis in Brazilian marsh deer (Blastocerus dichotomus).Vet Parasitol. 2006 Jun 30;139(1-3):262-6. doi: 10.1016/j.vetpar.2006.02.038. Epub 2006 Apr 18. Vet Parasitol. 2006. PMID: 16621285
-
Infections with Ehrlichia chaffeensis and Ehrlichia ewingii in persons coinfected with human immunodeficiency virus.Clin Infect Dis. 2001 Nov 1;33(9):1586-94. doi: 10.1086/323981. Epub 2001 Sep 24. Clin Infect Dis. 2001. PMID: 11568857 Review.
-
Current management of human granulocytic anaplasmosis, human monocytic ehrlichiosis and Ehrlichia ewingii ehrlichiosis.Expert Rev Anti Infect Ther. 2009 Aug;7(6):709-22. doi: 10.1586/eri.09.44. Expert Rev Anti Infect Ther. 2009. PMID: 19681699 Free PMC article. Review.
Cited by
-
Human monocytotropic ehrlichiosis-A systematic review and analysis of the literature.PLoS Negl Trop Dis. 2024 Aug 2;18(8):e0012377. doi: 10.1371/journal.pntd.0012377. eCollection 2024 Aug. PLoS Negl Trop Dis. 2024. PMID: 39093857 Free PMC article.
-
Global Prevalence of Anaplasma phagocytophilum in Cattle: A One Health Perspective, Meta-Analysis and Future Predictions (up to 2035).Vet Med Sci. 2025 Mar;11(2):e70251. doi: 10.1002/vms3.70251. Vet Med Sci. 2025. PMID: 39969156 Free PMC article. Review.
-
Challenges of Diagnosing Severe Ehrlichiosis in Orthotopic Liver Transplant Recipients.Case Rep Transplant. 2021 Nov 17;2021:8285326. doi: 10.1155/2021/8285326. eCollection 2021. Case Rep Transplant. 2021. PMID: 34840851 Free PMC article.
-
Donor-derived Ehrlichiosis: 2 Clusters Following Solid Organ Transplantation.Clin Infect Dis. 2022 Mar 9;74(5):918-923. doi: 10.1093/cid/ciab667. Clin Infect Dis. 2022. PMID: 34329411 Free PMC article.
-
Discovery and Surveillance of Tick-Borne Pathogens.J Med Entomol. 2021 Jul 16;58(4):1525-1535. doi: 10.1093/jme/tjaa269. J Med Entomol. 2021. PMID: 33313662 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials