title: Haplotype identification and detection of mitochondrial DNA heteroplasmy in Varroa destructor mites using ARMS and PCR-RFLP methods.
abstract: In the present study, amplification refractory mutation system (ARMS) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods were used for identification of recently described Serbia 1 (S1) and Peshter 1 (P1) mitochondrial haplotypes of Varroa destructor. Based on single nucleotide polymorphisms (SNPs) within cytochrome oxidase 1 (cox1) and cytochrome b (cytb) gene sequences, a total of 64 adult V. destructor females were analyzed from locations where the S1 and P1 haplotypes had been detected previously. Results of haplotype identification obtained by ARMS and PCR-RFLP methods were completely consistent with the sequencing data. Furthermore, in some analyzed samples the occurrence of site heteroplasmy at haplotype-defining sites was detected, as it was confirmed by double peaks in the sequence chromatograms. Neither mites with simultaneous nucleotide variability, nor those with combined SNP and heteroplasmy in cox1 and cytb were found. Given that this is the first occurrence of site heteroplasmy in V. destructor, the origin of this phenomenon and possible specific traits of heteroplasmic mites have yet to be determined.
authors.affiliation: Department of Parasitology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia. gajicb@vet.bg.ac.rs.
authors.forename: Bojan
authors.initials: B
authors.lastname: Gajić
authors.affiliation: Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia.
authors.forename: Jevrosima
authors.initials: J
authors.lastname: Stevanović
authors.affiliation: Laboratory for Medical Entomology, Department for Parasitology, Center of Exellence for Toxoplasmosis and Medical Entomology, Institute for Medical Research, University of Belgrade, Dr. Subotića 4, POB 102, Belgrade, 11129, Serbia.
authors.forename: Željko
authors.initials: Ž
authors.lastname: Radulović
authors.affiliation: Department of Parasitology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Kulišić
authors.affiliation: Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia.
authors.forename: Branislav
authors.initials: B
authors.lastname: Vejnović
authors.affiliation: Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia.
authors.forename: Uroš
authors.initials: U
authors.lastname: Glavinić
authors.affiliation: Department of Biology, Faculty of Veterinary Medicine, University of Belgrade, Bul. oslobodjenja 18, Belgrade, 11000, Serbia.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Stanimirović
mesh_terms: D000818:Animals; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D006239:Haplotypes; D021141:Nucleic Acid Amplification Techniques / Q000191:economics; D016133:Polymerase Chain Reaction / Q000191:economics; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D055771:Serbia; D057077:Varroidae / Q000235:genetics*
chemical_list: D004272:DNA, Mitochondrial
keywords: ARMS; Haplotype; PCR–RFLP; Varroa destructor; mtDNA heteroplasmy