Evaluation of the Effectiveness of Genetic Markers of Mycobacteria for Assessing the Disinfection Quality by Viability Real Time PCR

Khammadov, Nail I.; Александрова, Н. М.; Khammadova, Alfiya V.; Shuralev, Eduard A.

doi:10.1007/s12668-019-00654-8

Cited by 3 publications

(3 citation statements)

References 15 publications

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“…At the next stage, the species was confirmed by PCR. When creating a tool for the indication of microscopic fungi of the Fusarium genus by PCR, the following resources were used: National Center for Biotechnology Information resources ( https://www.ncbi.nlm.nih.gov ); “BLAST,” basic local alignment search tool utility ( https://blast.ncbi.nlm.nih.gov/Blast.cgi ); Vector NTI 9.1.0 program, bioinformatics analysis methodology is similar to our earlier papers [ 26 , 27 ]. The material for the research was the washings from the nutrient media of colonies of microscopic fungi.…”

Section: Methodsmentioning

confidence: 99%

A Case of Laying Hens Mycosis Caused by Fusarium proliferatum

Potekhina¹,

Tarasova²,

Matrosova³

et al. 2023

Veterinary Medicine International

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In this article, we present the first case report of a chicken mycosis caused by F. proliferatum occurred on a private farm in the Russian Federation. Lesions on the skin of the legs and scallops were reported. The object of this study was samples of feed and pathological material from sick hens-layers. Mycological analysis included determination of the total number of fungi (TNF) and identification and determination of the toxicity and pathogenicity of the isolates. The identification of the isolate was carried out taking into account direct microscopy, morphological features, and the method of molecular genetic analysis. Microscopic fungi of the genus Penicillium and Rhizopus were isolated by mycological analysis of the feed. The test feed was nontoxic. Mycological examination of pathological material (scrapings from the combs and affected legs) identified an isolate of Fusarium proliferatum, which showed toxicity on biological objects (protozoa, rabbits) and pathogenicity (white mice). Dermal application of F. proliferatum suspension was accompanied by reddening of the rabbit skin. Intraperitoneal injection of fungal spores caused mycosis in white mice. Polymerase chain reaction (PCR) made it possible to identify this type of microscopic fungus (F. proliferatum) with high accuracy in the samples under study. The research results allow us to consider F. proliferatum as a cause of poultry disease against the background of predisposing factors in the form of desquamation of the stratum corneum of the skin against the background of immunosuppression and metabolic disorders caused by an imbalance in the diet.

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Section: Methodsmentioning

confidence: 99%

A Case of Laying Hens Mycosis Caused by Fusarium proliferatum

Potekhina¹,

Tarasova²,

Matrosova³

et al. 2023

Veterinary Medicine International

Self Cite

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“…The methodology used in this work for analysis of the genomes of the seven FMDV serotypes (Table 1) were as described for other microorganisms [20,21]. The nucleotide sequences of the desired virus were determined by searching the National Center for Biotechnology Information (NCBI) resource databases.…”

Section: Methodsmentioning

confidence: 99%

“…PCR was performed on a C1000 amplifier with a CFX96 optical unit (BioRad). The methodology for PCR amplification is similar to that described previously [20], with the following modifications: probe for PCR, direct and reverse primers were developed in the framework of this work; the primer annealing temperature was 58.5°C; and the PCR (fluorescence) result was detected on each PCR cycle, at 58.5°C via Rox and Cy5 channels.…”

Section: Methodsmentioning

confidence: 99%

Genetic Polymorphisms of Foot-and-Mouth Disease Virus

Ildarovich¹

2021

Some RNA Viruses

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The aim of the work is to search for loci of the genome of various types of foot-and-mouth disease virus (FMDV), characterized by the lowest variability, for use as genetic markers in the polymerase chain reaction (PCR) of virus identification. The nucleotide sequences of the genomes of FMDV of types A, Asia-1, C, O, and SAT (1, 2, and 3) were analyzed. When aligning the genomes of isolates of each type of virus, potentially conservative sites were identified. Comparing these loci, different types of the virus have one, the most conserved locus. Subsequent basic local alignment search tool (BLAST) analysis established the correspondence of the conservative locus to the FMDV genome, and primers and a probe were developed to amplify this locus.

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Search for Genetic Markers of Tick-Borne Encephalitis Virus for Its Specific Indication, Genomic Analysis

Khammadov¹

2020

Problemy Osobo Opasnykh Infektsii

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Objective of the study is to analyze the genetic markers of tick-borne encephalitis virus, which can be used to specifically indicate the maximum number of virus strains and isolates.Materials and methods. Plasmid DNA and nucleic acids of the tick, the genus Ixodes and Dermacentor, were used as amplified material. Polymerase chain reaction (PCR) was conducted on C1000 amplifier with a CFX96 optical unit (BioRad). The species (strain) variety of detected organisms, using the analyzed genetic markers, was determined in the nBLAST software utility. The design of the nucleotide sequences of primers and probes was performed using “Vector NTI 9.1.0” (Invitrogen Corporation). Nucleic acids were isolated by magnetic sorption with a reagent panel MAGNO-sorb. Primers and probes were synthesized at “Evrogen” company, Moscow, Russia. The reagents for PCR were manufactured by “Syntol” company, Moscow, Russia.Results and discussion. When indicating the genome of the tick-borne encephalitis virus, the main criterion for choosing a marker sequence is the specific detection of nucleic acids of only the desired microorganism. The genomewide nucleotide sequences of various strains and isolates of tick-borne encephalitis virus were analyzed to search for a specific marker nucleotide sequence. Mutual comparison of all the above mentioned genomes made it possible to determine 7 conventionally conservative loci characterized by minimal nucleotide variability. The further work was based on the results of alignment of the nucleotide sequences of tick-borne encephalitis virus isolates, including the nucleotide sequences of heterogeneous microorganisms; primers and probes were designed to amplify each of the marker loci; the most analytically significant oligonucleotides were developed based on loci 4 and 7. Amplification with oligonucleotide primers to indicate tick-borne encephalitis virus was effective, both in a separate PCR with a positive control, and in combination with tick DNA.

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Evaluation of the Effectiveness of Genetic Markers of Mycobacteria for Assessing the Disinfection Quality by Viability Real Time PCR

Cited by 3 publications

References 15 publications

A Case of Laying Hens Mycosis Caused by Fusarium proliferatum

A Case of Laying Hens Mycosis Caused by Fusarium proliferatum

Genetic Polymorphisms of Foot-and-Mouth Disease Virus

Search for Genetic Markers of Tick-Borne Encephalitis Virus for Its Specific Indication, Genomic Analysis

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