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[THE HIGHLY EFFECTIVE DETECTION OF DNA RICKETTSIA USING TECHNIQUE OF POLYMERASE CHAIN REACTION IN REAL-TIME].
- Source :
-
Klinicheskaia laboratornaia diagnostika [Klin Lab Diagn] 2015 Dec; Vol. 60 (12), pp. 39-43. - Publication Year :
- 2015
-
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.
- Subjects :
- Animals
DNA Primers chemical synthesis
DNA Primers chemistry
DNA Probes chemical synthesis
DNA Probes chemistry
Gene Expression
Ixodes chemistry
Rickettsia classification
Rickettsia isolation & purification
Russia
Sensitivity and Specificity
Bacterial Proteins genetics
Citrate (si)-Synthase genetics
DNA, Bacterial genetics
Ixodes microbiology
Real-Time Polymerase Chain Reaction methods
Rickettsia genetics
Subjects
Details
- Language :
- Russian
- ISSN :
- 0869-2084
- Volume :
- 60
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Klinicheskaia laboratornaia diagnostika
- Publication Type :
- Academic Journal
- Accession number :
- 27032252