title: [THE HIGHLY EFFECTIVE DETECTION OF DNA RICKETTSIA USING TECHNIQUE OF POLYMERASE CHAIN REACTION IN REAL-TIME].
abstract: The article considers development of highly effective technique of detection of genetic material of ricketsia based on polymerase chain reaction in real-time using original primers to the most conservative sites of gene of citrate synthase (gItA). The analytical sensitivity of the developed polymerase chain reaction in real-time test permits to detect from 80 genome equivalents in analyzed sample during three hours. The high specificity of test-system is substantiated by detection of nucleotide sequences of amplificated fragments of gene gltA. The approbation ofthe polymerase chain reaction in real-time test is carried out on collection of 310 ticks of species I. persulcatus, I. pavlovskyi, D. reticulatus. It is demonstrated that the developed alternate ofprimers and probe permits with high degree of sensitivity and specifcity to detect DNA of different species of ricketsia widespread on territory of Russia (R. sibirica, R. raoultii, R. helvetica, R. tarasevichiae). The proposed polymerase chain reaction in real-time test can be appliedfor isolation of fragment of gene gltA with purpose for detecting nucleotide sequence and subsequent genetic typing of ricketsia. The application ofthe proposed technique can facilitate task of monitoring hot spots of ricketsiosis.
authors.forename: M Yu
authors.initials: MY
authors.lastname: Kartashov
authors.forename: T P
authors.initials: TP
authors.lastname: Mikryukova
authors.forename: V A
authors.initials: VA
authors.lastname: Ternovoi
authors.forename: N S
authors.initials: NS
authors.lastname: Moskvitina
authors.forename: V B
authors.initials: VB
authors.lastname: Loktev
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002950:Citrate (si)-Synthase / Q000235:genetics*; D017931:DNA Primers / Q000138:chemical synthesis / Q000737:chemistry; D015342:DNA Probes / Q000138:chemical synthesis / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics*; D015870:Gene Expression; D018884:Ixodes / Q000737:chemistry / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012281:Rickettsia / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012426:Russia; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D002950:Citrate (si)-Synthase