期刊名称：Annals of Agricultural and Environmental Medicine
印刷版ISSN：1232-1966
电子版ISSN：1898-2263
出版年度：2015
卷号：22
期号：4
页码：642-646
DOI：10.5604/12321966.1185767
出版社：Institute of Agricultural Medicine in Lublin
摘要：Introduction. Ticks transmit a great variety of pathogenic microorganisms to humans and animals. The detection of tickbornepathogens (TBP) is mainly by molecular techniques based on polymerase chain reactions (PCR).Objective. To design and evaluate a multiplex PCR for the molecular screening of zoonotic TBP for exploratory studies.Material and methods. Control DNA from reference strains, DNA from experimentally-infected biological specimens,and from Rhipicephalus sanguineus ticks collected from domestic and homeless dogs were used. A multiplex PCR assay todetect the presence of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. was designed and optimized usingprimers previously reported for B. burgdorferi sensu lato and Anaplasma spp., while for Babesia spp. they were designed insilico. The multiplex PCR was evaluated on the DNA from biological samples.Results. A new set of specific primers for Babesia spp. was designed. Adjustment of the master mix reactive concentrationsand amplification conditions for the multiplex PCR allowed the successful amplification of the specific amplicons for eachmicrobial group from the control DNA and experimentally-infected biological specimens. The efficiency of the multiplexPCR amplifying three DNA targets was confirmed. Individual and co-infection of Anaplasma spp. and Babesia spp. weredetected in the R. sanguineus ticks from dogs.Conclusions. A multiplex PCR assay for the screening of three TBP is available. By using it, B. burgdorferi sensu lato, Anaplasmaspp. and Babesia spp. can be detected accurately in one PCR reaction
