Validation of PCR tests with Russian kits for the detection of Ralstonia solanacearum sensu lato in plant extracts
https://doi.org/10.69536/j3225-4992-5329-t
Abstract
The validation of two tests based on qPCR and a FLASH test with commercial kits for DNA isolation and amplification produced by Russian companies AgroDiagnostika and Syntol was assessed, as well as using Russian amplifiers for identification of the genetic material of a complex of causative agents of the quarantine disease, brown rot and bacterial wilt Ralstonia solanacearum sen su lato, in extract of potatoes and roses. The analytical sensitivity of tests for most strains was 102–103 CFU/ml. The repeatability and reproducibility of RT-PCR tests depended on the instrument used. The values of the working criteria up to 100% were obtained on a qPCR machine DT-light manufactured by DNA-Technology. Based on the test results, Agro-Diagnostika kits can be recommended for carrying out universal screening tests during laboratory studies of regulated products samples to identify pathogens of the species complex. The test with the Syntol kit, specific to R. solanacearum race 3 biovar 2, can be used to confirm a positive result of the screening test or as a screening test when examining samples from the spread areas of R. solanacearum race 3 biovar 2, as well as for differential diagnosis this group.
About the Authors
N. V. Drenova
FGBU “All-Russian Plant Quarantine Center” (FGBU “VNIIKR”)
Russian Federation
Russian Federation
Nataliya Drenova, Senior Researcher, Research and Methodology Department of Virology and Bacteriology, FGBU “VNIIKR”
Bykovo, Ramenskoye, Moscow Oblast
I. M. Ignatieva
FGBU “All-Russian Plant Quarantine Center” (FGBU “VNIIKR”)
Russian Federation
Russian Federation
Irina Ignatieva, Researcher, Bacteriology and GMO Analysis Laboratory, FGBU “VNIIKR”
Bykovo, Ramenskoye, Moscow Oblast
M. O. Kondratiev
FGBU “All-Russian Plant Quarantine Center” (FGBU “VNIIKR”)
Russian Federation
Russian Federation
Maksim Kondratiev, Agronomist, Research and Methodology Department of Virology and Bacteriology, FGBU “VNIIKR”
Bykovo, Ramenskoye, Moscow Oblast
Ye. Yu. Shneyder
FGBU “All-Russian Plant Quarantine Center” (FGBU “VNIIKR”)
Russian Federation
Russian Federation
Elena Shneyder, Senior Researcher, Research and Methodology Department of Virology and Bacteriology, FGBU “VNIIKR”
Bykovo, Ramenskoye, Moscow Oblast
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Review
For citations:
Drenova N.V., Ignatieva I.M., Kondratiev M.O., Shneyder Ye.Yu. Validation of PCR tests with Russian kits for the detection of Ralstonia solanacearum sensu lato in plant extracts. Plant Health and Quarantine. 2022;(1):2-20. https://doi.org/10.69536/j3225-4992-5329-t
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