Detection of Mixed Infections with "Candidatus Mycoplasma Haemominutum" and Mycoplasma Haemofelis Using Real-time Taqman Polymerase Chain Reaction
Correspondence: 1 Corresponding Author: Jane E Sykes, VM, Medicine and Epidemiology, 2108 Tupper Hall, University of California, Davis, Davis,, CA 95616, e-mail: jesykes{at}ucdavis.edu The objective of this study was to evaluate the use of real-time TaqMan PCR assays for detection of coinfections wi...
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Veröffentlicht in: | Journal of veterinary diagnostic investigation 2007-05, Vol.19 (3), p.250-255 |
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Zusammenfassung: | Correspondence: 1 Corresponding Author: Jane E Sykes, VM, Medicine and Epidemiology, 2108 Tupper Hall, University of California, Davis, Davis,, CA 95616, e-mail: jesykes{at}ucdavis.edu
The objective of this study was to evaluate the use of real-time TaqMan PCR assays for detection of coinfections with " Candidatus Mycoplasma haemominutum" (Mhm), and Mycoplasma haemofelis (Mhf), in vitro and over time in experimentally infected cats. First, the ability of each real-time PCR assay to detect and quantify mixed infections was determined in vitro by testing mixtures of plasmids containing Mhm and Mhf 16S rDNA with each assay. Subsequently, 4 specific pathogen-free (SPF) cats, 2 of which were splenectomized, were inoculated with blood from a cat infected with both Mhm and Mhf. Sixteen blood samples were then collected from each cat over a 55-day period. Each of the 64 postinoculation samples was tested using both conventional polymerase chain reaction (cPCR) and real-time PCR for the 16S rRNA gene of each organism. When applied to mixtures of plasmid DNA from each species, the results of quantitation with each of the real-time PCR assays approximately reflected the number of plasmid copies present. Forty-nine of 64 post-inoculation samples (77%) were positive using both cPCR and real-time PCR, 4 (6%) were positive using cPCR only, and 3 (5%) were positive using real-time PCR only. Both organisms were detected in 23 samples using real-time PCR. Mixed infections were not detected using cPCR. The size of the corresponding cPCR products suggested infection with Mhm in 4 and Mhf in 18 of these samples. The use of multiple separate real-time PCR assays rather than cPCR alone should thus be considered for epidemiologic studies of hemoplasmosis in cats.
Key Words: Anemia Haemobartonella hemoplasma quantitative PCR splenectomy |
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ISSN: | 1040-6387 1943-4936 |
DOI: | 10.1177/104063870701900304 |