METHOD FOR INDICATION AND IDENTIFICATION OF STRAINS OF THE PLAGUE PATHOGEN BY THEIR BELONGING TO THE SPECIES YERSINIA PESTIS, TO SUBSPECIES, BIOVARS, PHYLOGENETIC BRANCHES AND BY THE PRESENCE OF GENES OF PATHOGENICITY MAIN FACTORS BY THE DNA-CHIP METHOD Russian patent published in 2020 - IPC C12Q1/68 

Abstract RU 2734636 C1

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to indication and identification of plague agent strains by their belonging to the species Yersinia pestis, to subspecies, biovars, phylogenetic branches by presence of genes of pathogenicity main factors by DNA-chip method. Method involves recovering a DNA strain, carrying out multiplex PCR in one step in six reaction mixtures on target DNA: ‘pla’, ‘3a’, ‘cafl’, ‘yihN’ in first; ‘irp2’, ‘lcrV’ in second; ‘glpD’, ‘Pro’, ‘Med24’, ‘45’ in third; ‘Med70’, ‘Phage’ in fourth; ‘Alt’, ‘His’, ‘U1’ in fifth; ‘Caucas’, ‘Tal’, ‘Micr’ in sixth with subsequent hybridisation with immobilized probes, indication and identification of strains based on the presence and absence of fluorescence signals. Fluorescence on targets "yihN" and "3a" will be observed in all strains of Y. pestis. If differentiating subspecies, there will be no fluorescence signal in the Y. pestis strains of the main subspecies on target "45", in strains of Altaic biovar of Central Asian sub-species by target "Alt", in Gissar biovar of Central Asian subspecies by "His" target, in Talas biovar of Central Asian sub-species by target "Ta1", in microtias biovar of Central Asian subspecies by target "Micr", in ulegeica subspecies strains by target "U1", strains of Caucasian subspecies by "Caucas" target. If differentiating the Y. pestis strains of the main subspecies as belonging to the biovars, there will be no fluorescence signal in the strains of Oriental biovar (1.ORI) on the "glpD" target, in all strains of medieval biovar except 2.MED0 on target "Med24" and fluorescence signals in strains of branch 2.MED0 on target "pCKP", in strains of antique biovar of the main subspecies on targets "glpD" and "Med24" will be noted. Phylogenetic branch "1" (1.ANT and 1.ORI) strains will be characterized by presence of fluorescent signal on "Phage" target, phylogenetic branch "2" (2.ANT and 2.MED) strains will be characterized by absence of fluorescence signal on "Med70" target. When detecting the main pathogenicity genes, there will be a fluorescence signal in Y. pestis strains containing an HPI high pathogenicity island in the chromosome pigmentation region of the target "irp2", in strains carrying pla gene of plasmid pPst on target "pla", in strains with gene lcrV of plasmid pCad on target "lcrV", in strains containing cafl gene of plasmid pFra on target "caf".

EFFECT: invention makes it possible to determine affiliation to species, subspecies, biovars, phylogenetic branches and presence of main genes of pathogenicity by DNA-chip method, which provides isolation of DNA-strain, conducting six multiplex PCR on target DNA ‘yihN’, ‘3a’, ‘45’, ‘Alt’, ‘His’, ‘U1’, ‘Caucas’, ‘Tal’, ‘Micr’, ‘glpD’, ‘Med24’, ‘Pro’, ‘Med70’, ‘Phage’, ‘pla’, ‘cafl’, ‘irp2’, ‘lcrV’, with subsequent hybridisation with immobilized probes, indication and identification of strains based on the presence and absence of fluorescence signals in accordance with the tables.

1 cl, 6 tbl, 9 ex

Similar patents RU2734636C1

Title Year Author Number
METHOD FOR IDENTIFICATION OF YERSINIA PESTIS STRAINS OF A MEDIEVAL BIOVAR WITH SUBSEQUENT DIFFERENTIATION BY PHYLOGENETIC AFFILIATION BY POLYMERASE CHAIN REACTION WITH HYBRIDIZATION-FLUORESCENT ACCOUNT OF RESULTS 2018
  • Nosov Nikita Yurevich
  • Nikiforov Konstantin Alekseevich
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2705813C1
SET AND METHOD FOR ACCELERATED PLAGUE MICROBE IDENTIFICATION AND SIMULTANEOUS DIFFERENTIATION OF VIRULENT AND AVIRULENT STRAINS OF YPESTIS BY PLASMID PROFILING THEREOF 2011
  • Kuklev Vasilij Evgen'Evich
  • Osina Natal'Ja Aleksandrovna
  • Bugorkova Tat'Jana Vasil'Evna
  • Kutyrev Vladimir Viktorovich
RU2473701C1
BACTERIAL STRAINS KIT USED FOR TRAINING ON MICROBIOLOGY AND PLAGUE LABORATORY DIAGNOSTICS METHODS 2016
  • Sazanova Elena Vladimirovna
  • Malyukova Tatyana Anatolevna
  • Popov Yurij Alekseevich
  • Kuklev Vasilij Evgenevich
  • Malakhaeva Alina Nikolaevna
  • Kutyrev Vladimir Viktorovich
RU2642322C1
DIFFERENTIATION METHOD OF BIOVARS AND GENOVARIATIONS OF Yersinia pestis STRAINS OF MAIN SUBSPECIES BY MEANS OF POLYMERASE CHAIN REACTION 2014
  • Nikiforov Konstantin Alekseevich
  • Odinokov Georgij Nikolaevich
  • Oglodin Evgenij Gennad'Evich
  • Kukleva Ljubov' Mikhajlovna
  • Shavina Natal'Ja Jur'Evna
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2565554C2
METHOD FOR IDENTIFYING YERSINIA PESTIS AND YERSINIA PSEUDOTUBERCULOSIS AND SIMULTANEOUS DIFFERENTIATION OF YERSINIA PESTIS OF MAIN AND CENTRAL ASIAN SUBSPECIES BY MULTIPLEX PCR 2020
  • Morozov Oleg Alekseevich
  • Nikiforov Konstantin Alekseevich
  • Oglodin Evgenij Gennadevich
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2737775C1
METHOD OF DIFFERENTIATING TYPICAL AND ATYPICAL STRAINS Yersinia pestis OF MEDIEVAL BIOVAR BY PCR METHOD WITH HYBRIDISATION-FLUORESCENT RECORDING RESULTS 2014
  • Oglodin Evgenij Gennad'Evich
  • Nikiforov Konstantin Alekseevich
  • Odinokov Georgij Nikolaevich
  • Kukleva Ljubov' Mikhajlovna
  • Anisimova Ljubov' Vladimirovna
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2550257C2
METHOD FOR DIFFERENTIATING YERSINIA PESTIS STRAINS OF VARIOUS SUBSPECIES AND BIOVARS BY POLYMERASE CHAIN REACTION AND MULTILOCUS SEQUENCE TYPING 2011
  • Eroshenko Galina Aleksandrovna
  • Kukleva Ljubov' Mikhajlovna
  • Pavlova Alla Ivanovna
  • Odinokov Georgij Nikolaevich
  • Krasnov Jaroslav Mikhajlovich
  • Guseva Natal'Ja Petrovna
  • Kutyrev Vladimir Viktorovich
RU2471872C1
METHOD OF DETERMINING THE PHYLOGENETIC AFFILIATION OF STRAINS OF YERSINIA PESTIS OF THE MAIN SUBSPECIES BY ALLELE-SPECIFIC PCR IN REAL TIME 2022
  • Nikiforov Konstantin Alekseevich
  • Oglodin Evgenij Gennadevich
  • Kukleva Lyubov Mikhajlovna
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2799415C1
METHOD OF DIFFERENTIATION OF YERSINIA PESTIS STRAINS OF MEDIEVAL BIOVAR ACCORDING TO PHYLOGENETIC AFFILIATION WITH PCR METHOD WITH ELECTROPHORETIC ACCOUNTING OF RESULTS 2018
  • Nosov Nikita Yurevich
  • Oglodin Evgenij Gennadevich
  • Nikiforov Konstantin Alekseevich
  • Kukleva Lyubov Mikhajlovna
  • Eroshenko Galina Aleksandrovna
  • Kutyrev Vladimir Viktorovich
RU2684231C1
METHOD OF IDENTIFICATION AND INTRAGENERIC DIFFERENTIATION OF STRAINS OF SPECIES YERSINIA PESTIS 2009
  • Trukhachev Aleksej Leonidovich
  • Lebedeva Svetlana Aleksandrovna
  • Vasil'Eva Ekaterina Anatol'Evna
  • Ivanova Vera Stepanovna
  • Rakin Aleksandr Vladimirovich
RU2404251C1

RU 2 734 636 C1

Authors

Nikiforov Konstantin Alekseevich

Utkin Denis Valerevich

Kukleva Lyubov Mikhajlovna

Eroshenko Galina Aleksandrovna

Kutyrev Vladimir Viktorovich

Dates

2020-10-21Published

2020-02-12Filed