Multiplex PCR for detection of Anaplasma marginale, A. bovis and A. platys in cattle.

Autor: Teja MMS; Department of Veterinary Parasitology, Veterinary College, Bengaluru, KVAFSU (BIDAR), Bidar, India., Mamatha GS; Department of Veterinary Parasitology, Veterinary College, Bengaluru, KVAFSU (BIDAR), Bidar, India., Lakkundi JN; Department of Veterinary Parasitology, Veterinary College, Bengaluru, KVAFSU (BIDAR), Bidar, India., Chandranaik BM; Department of Biological Productions, IAH&VB, Bengaluru, KVAFSU (BIDAR), Bidar, India., Murthy CMK; Department of Veterinary Parasitology, Veterinary College, Bengaluru, KVAFSU (BIDAR), Bidar, India.; Department of Veterinary Parasitology, Veterinary College, Bidar, KVAFSU (BIDAR), Bidar, India., Gomes AR; Department of Biological Productions, IAH&VB, Bengaluru, KVAFSU (BIDAR), Bidar, India.
Jazyk: angličtina
Zdroj: Journal of parasitic diseases : official organ of the Indian Society for Parasitology [J Parasit Dis] 2023 Sep; Vol. 47 (3), pp. 659-663. Date of Electronic Publication: 2023 Jun 23.
DOI: 10.1007/s12639-023-01606-6
Abstrakt: Anaplasma genus has been classified under the family Anaplasmataceae of order Rickettsiales, which contains the spp. Anaplasma marginale , Anaplasma bovis, Anaplasma centrale , Anaplasma platys and Anaplasma phagocytophilum is accountable for a broad range of diseases in both man and animal medicine around the globe. A multiplex PCR procedure was designed to identify A. marginale , A. bovis , and A. platys simultaneously by employing species-specific primers targeting the msp 4 ( Anaplasma marginale ), 16S rRNA ( Anaplasma bovis ), and groEL ( Anaplasma platys ) genes. The sensitivity of the assay was ascertained by tenfold dilutions of DNA obtained from cattle blood infected with A. marginale , A. bovis , and A. platys was used to evaluate the test sensitivity. A total of 31 Anaplasma genus positive samples were subjected to mPCR by using species specific primers for simultaneous detection of all the three species. All the 31 samples showed amplification of 753 bp fragment specific for A. marginale and 61.29% of the samples showed amplification of 547 bp fragment specific for A. bovis . The 470 bp DNA fragment specific for A. platys was found in 96.7% of samples. The lower limit of detection of the method for A. platys, A. marginale and A. bovis was found to be 4.4, 0.44 and 0.044 ng/µl, respectively.
Competing Interests: Conflict of interestAuthors have no conflict of interest.
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Databáze: MEDLINE
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