Duplex TaqMan real-time PCR assay for simultaneous detection and quantification of Anaplasma capra and Anaplasma phagocytophilum infection

This study aimed to develop a rapid, sensitive and reliable duplex real-time PCR assay for the specific detection and differentiation of these Anaplasma species.We designed primers and probes against the conserved regions of A. capra groEL and A. phagocytophilum 16S rRNA genes. A range of PCR-related parameters were evaluated such as the dosage of primers and probes, and annealing temperature. The specificity, sensitivity and repeatability of this assay were evaluated. Assay performance was further evaluated using samples collected from 124 goats in four regions of Henan, China. This set of samples was also tested using conventional PCR under conditions previously described.The developed duplex real-time PCR assay allowed the simultaneous detection of A. capra and A. phagocytophilum in a reasonably short time at levels as small as 102 copies/μL, respectively, with optimal specificity and reproducibility. In addition, this duplex real-time PCR assay is the first DNA-based method designed to detect A. capra and A. phagocytophilum, and will be valuable for timely diagnosis and treatment of these infections.
Source: Molecular and Cellular Probes - Category: Molecular Biology Source Type: research

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Source: International Journal for Parasitology: Parasites and Wildlife - Category: Parasitology Source Type: research
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Source: Frontiers in Microbiology - Category: Microbiology Source Type: research
Vector-Borne and Zoonotic Diseases, Ahead of Print.
Source: Vector-Borne and Zoonotic Diseases - Category: Infectious Diseases Authors: Source Type: research
Vector-Borne and Zoonotic Diseases, Ahead of Print.
Source: Vector-Borne and Zoonotic Diseases - Category: Infectious Diseases Authors: Source Type: research
Vector-Borne and Zoonotic Diseases, Ahead of Print.
Source: Vector-Borne and Zoonotic Diseases - Category: Infectious Diseases Authors: Source Type: research
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Source: Ticks and Tick borne Diseases - Category: Zoology Source Type: research
J. R. Head et al.
Source: Emerging Infectious Diseases Journal - Category: Infectious Diseases Source Type: research
This study assessed the prevalence, risk factors, and co-infectivity of CTBP such as piroplasms, rickettsiae, Coxiella burnetii, hepatozoa, hemotropic mycoplasmas, and Borrelia spp., using PCR and phylogenetic analyses. Of the 510 dogs tested, three CTBP, including Anaplasma phagocytophilum 16S rRNA (13; 2.6%), Hepatozoon canis 18S rRNA (2; 0.4%), and Mycoplasma haemocanis 16S rRNA (1; 0.2%) were detected. PCR adapted to amplify A. phagocytophilum msp2 and groEL genes generated amplicons thereof in two out of 13 positive dogs. One shelter dog was co-infected with A. phagocytophilum and M. haemocanis. However, Ehrlichia spp...
Source: Ticks and Tick borne Diseases - Category: Zoology Source Type: research
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Source: Ticks and Tick borne Diseases - Category: Zoology Source Type: research
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Source: Ticks and Tick borne Diseases - Category: Zoology Source Type: research
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