scholarly journals Molecular genetic analysis of the strain Leningrad-16 used for the production of measles vaccine

2020 ◽  
Vol 97 (2) ◽  
pp. 182-189
Author(s):  
Georgy M. Ignatyev ◽  
Alena V. Atrasheuskaya ◽  
Lidiya L. Sukhanova ◽  
Elena S. Sidorenko ◽  
Nina A. Netesova
Keyword(s):  
Vaccine Strain ◽  
Measles Virus ◽  
Restriction Analysis ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Rt Pcr ◽  
Virus Genotype ◽  
Molecular Genetic Study ◽  
Production Stages ◽  
Pcr Method

Aim. To study the genetic stability of the measles virus strain Leningrad-16 (L-16) used for the production of vaccine at JSC NPO Mikrogen.Materials and methods. A series of production and sowing strains of L-16 (JSC NPO Mikrogen), ready-made series of measles vaccines from various manufacturers, and the strain of measles virus genotype D6 were studied. Molecular genetic study of the strains was performed using RT-PCR followed by restriction analysis and sequencing.Results. The complete genome sequences of the production and sowing strains of L-16 that are used for vaccine production were obtained. The sequence of the vaccine strain was deposited in GenBank. Strain L-16 was confirmed to be genetically stable. The obtained data demonstrated the possibility of using the RT-PCR method with subsequent restriction analysis to confirm the authenticity of the vaccine strain L-16 in finished mono and three component vaccines.Conclusion. The results of the study suggest the applicability of the molecular genetic methods to confirm the authenticity of the studied strains not only at the production stages, but also in the finished series of vaccines.

scholarly journals Molecular-genetic study of the RA-27/3 strain used for production of rubella vaccine

2019 ◽  
pp. 38-46 ◽  
Author(s):  
G. M. Ignatev ◽  
E. V. Atrashevskaya ◽  
L. L. Suchanova ◽  
E. S. Sidorenko ◽  
N. A. Netesova
Keyword(s):  
Virus Strain ◽  
Rubella Virus ◽  
Genetic Study ◽  
Molecular Genetic ◽  
Vaccine Virus ◽  
Rt Pcr ◽  
Molecular Genetic Study ◽  
Virus Identification ◽  
Production Stages ◽  
Combined Vaccine

Aim. In order to study rubella virus strain RA-27/3 genetic stability, used for the vaccines production, a molecular genetic study was conducted. Materials and methods. In the study different series of master and work seed of RA-27/3 rubella virus strain by «Microgen», a few lots of rubella vaccines by the different manufacturers, as well as strain «Orlov» of rubella virus were used. RT-PCR followed by restriction, sequencing were performed . Results. Full-genomic sequences of the rubella virus strain RA-27/3 by «Microgen», were obtained and presented to GenBank. The full structure correspondence of RA-27/3 rubella virus strain by «Microgen» to the similar rubella strains used by GSK and Merck & Co Inc. has been shown. The RT-PCR method with the subsequent restriction was fulfilled using only domestic reagents. The developed method has been demonstrated as applicable for the identification of the RA-27/3 rubella virus strain as in monopreparation as well as in the combined vaccine preparation. Conclusion. The data obtained make it possible to suggest application of the molecular genetic methods for the vaccine virus identification not only at the production stages, but also in the finished vaccine lots.

scholarly journals Greek measles epidemic strain, 2005–2006

2006 ◽  
Vol 135 (4) ◽  
pp. 570-573 ◽  
Author(s):  
G. GIOULA ◽  
A. PAPA ◽  
M. EXINDARI ◽  
A. MELIDOU ◽  
D. CHATZIDIMITRIOU ◽  
...  
Keyword(s):  
Phylogenetic Tree ◽  
Vaccine Strain ◽  
Virus Strain ◽  
Measles Virus ◽  
Molecular Study ◽  
N Gene ◽  
Epidemic Strain ◽  
Rt Pcr ◽  
Measles Outbreak ◽  
Genotype A

SUMMARYThe purpose of this work was the molecular study of the virus strain that caused the last measles outbreak in Greece. Twenty-four saliva specimens were obtained from selected patients serologically confirmed as measles cases between December 2005 and March 2006. Measles virus (MV) detection was performed by a nested RT–PCR. The 447-bp segment of the N gene of these MV strains was used for genotyping. The N gene sequences of the Greek MV strains were identical to each other, therefore a phylogenetic tree was constructed using one representative MV (ThesGRE/06). Our data show that the MV strain which caused the 2005–2006 outbreak in Greece belongs to genotype D6, and differs by 0·68% from the New Jersey D6 strain and by 5·5% from the MV vaccine strain Edmonston B (U03656) belonging to genotype A.

scholarly journals Rapid Identification of Measles Virus Vaccine Genotype by Real-Time PCR

2016 ◽  
Vol 55 (3) ◽  
pp. 735-743 ◽  
Author(s):  
Felicia Roy ◽  
Lillian Mendoza ◽  
Joanne Hiebert ◽  
Rebecca J. McNall ◽  
Bettina Bankamp ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Measles Virus ◽  
Rapid Identification ◽  
Rt Pcr ◽  
Outbreak Response ◽  
Reverse Transcription Pcr ◽  
Pcr Method ◽  
Measles Outbreaks ◽  
Genotype A

ABSTRACT During measles outbreaks, it is important to be able to rapidly distinguish between measles cases and vaccine reactions to avoid unnecessary outbreak response measures such as case isolation and contact investigations. We have developed a real-time reverse transcription-PCR (RT-PCR) method specific for genotype A measles virus (MeV) (MeVA RT-quantitative PCR [RT-qPCR]) that can identify measles vaccine strains rapidly, with high throughput, and without the need for sequencing to determine the genotype. We have evaluated the method independently in three measles reference laboratories using two platforms, the Roche LightCycler 480 system and the Applied Biosystems (ABI) 7500 real-time PCR system. In comparison to the standard real-time RT-PCR method, the MeVA RT-qPCR showed 99.5% specificity for genotype A and 94% sensitivity for both platforms. The new assay was able to detect RNA from five currently used vaccine strains, AIK-C, CAM-70, Edmonston-Zagreb, Moraten, and Shanghai-191. The MeVA RT-qPCR assay has been used successfully for measles surveillance in reference laboratories, and it could be readily deployed to national and subnational laboratories on a wide scale.

scholarly journals Molecular genetic analysis of some North African barley germplasms

2017 ◽  
Vol 109 (2) ◽  
pp. 187
Author(s):  
Reda Gaafar ◽  
Mai Allam ◽  
Rasha Sabry ◽  
Mahmoud Saker
Keyword(s):  
Rapd Markers ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
North African ◽  
Rt Pcr ◽  
Breeding Selection ◽  
High Level ◽  
Barley Genotypes ◽  
Selection Of ◽  
Selection Of Resistance

<p>Isozyme and RAPD markers were used to characterize 29 barley accessions, which were collected from North Africa. In addition, resistance gene sequences were employed to develop molecular markers using RT-PCR approach. High level of polymorphism was found with both RAPD and isozyme markers, where RAPD showed that 60 % of amplified bands were polymorphic. Peroxidase showed three polymorphic loci (7 allelic bands). Isozymes cluster analysis successfully separated the barley accessions into three geographically distinct groups. RAPD investigation demonstrated that Egyptian accessions were grouped into two obvious groups. Moreover, the Tunisian accessions showed no distinct clustering, while high dissimilarities were revealed by the Algerian accessions. In the RT-PCR, from six primer pairs selected, primer pair AF092524P1P2 successfully amplified two specific amplicons of approximately (340 &amp; 220 bp) and (360 &amp; 270 bp), respectively in two Egyptian barley genotypes (El-Awamah and Awlad-Ali). One primer pair DN988165P1P2 gave only one specific amplicon in both barley genotypes of 250 and 270 bp, respectively. The markers developed could be used in improving barley crop by assisting in breeding selection of resistance genotypes.</p>

scholarly journals Molecular Genetic Testing of Stability and Identification of Vnukovo-32 Strain Used for Production of the Cultural Concentrated Purified Inactivated Dry Rabies Vaccine

2020 ◽  
Vol 20 (2) ◽  
pp. 107-115
Author(s):  
G. M. Ignatyev ◽  
A. S. Oksanich ◽  
L. P. Antonova ◽  
T. G. Samartseva ◽  
S. V. Mosolova ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Rabies Virus ◽  
Dosage Form ◽  
Restriction Analysis ◽  
Molecular Genetic ◽  
Rabies Vaccine ◽  
Strain Identification ◽  
Production Stages ◽  
Rabies Virus Strain ◽  
Production Strain

Rabies is an acute viral disease caused by a virus of the Rhabdoviridae family of the Lyssavirus genus, which affects the central nervous system and is characterised by absolute mortality. Vaccination is the only way to prevent the disease in humans. One of the products used for vaccination is a cultural concentrated purified inactivated dry rabies vaccine produced by the Federal State Budgetary Institution of Science “Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences” (hereinafter—Chumakov Center).The aim of the study was to examine the structure of the working virus seed of Vnukovo-32 strain used by the Chumakov Center for rabies vaccine production, to assess its genetic stability during production, to explore the possibility of using molecular genetic methods for identification of the production strain in the finished dosage form, and to study the nucleotide sequence of the CVS strain.Materials and methods: Vnukovo-32 rabies virus production strain, working virus seeds, finished batches of the rabies vaccine, CVS fixed rabies virus strain used in the assessment of specific immunity. The molecular genetic study was performed using RT-PCR followed by restriction and sequencing.Results: the paper presents the results of nucleotide sequence analysis of the G gene fragment obtained from the Vnukovo-32 production strain, batches of the working virus seed, and finished batches of the rabies vaccine produced in 2012, 2018, and 2019, and the CVS fixed rabies virus strain used in the assessment of the vaccine’s specific immunity. The study demonstrated that restriction analysis could be used for Vnukovo-32 strain identification at all production stages, including the finished dosage form.Conclusion: Vnukovo-32 and CVS strains used by the Chumakov Center are rabies viruses. Analysis of the nucleotide sequence of the G gene fragment showed that the Vnukovo-32 strain remains stable throughout different production stages. The obtained nucleotide sequence of gene G of the Vnukovo-32 strain was deposited in GenBank (accession number MN116503). The study demonstrated that restriction analysis could be used for Vnukovo-32 strain identification at all production stages, including the finished dosage form. 

scholarly journals Primary synovial sarcoma of the heart: a cytogenetic and molecular genetic analysis combining RT-PCR and COBRA-FISH of a case with a complex karyotype

2004 ◽  
Vol 17 (11) ◽  
pp. 1434-1439 ◽  
Author(s):  
Hans Martin Hazelbag ◽  
Károly Szuhai ◽  
Hans J Tanke ◽  
Carla Rosenberg ◽  
Pancras C W Hogendoorn
Keyword(s):  
Genetic Analysis ◽  
Synovial Sarcoma ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Complex Karyotype ◽  
Rt Pcr

scholarly journals THE MOLECULAR-GENETIC ANALYSIS OF POLYMORPHISM OF IL-4, TNF-α GENES IN PATIENTS WITH LIVER CIRROSIS AND PORTAL HYPERTENSION

2017 ◽  
pp. 110-114
Author(s):  
A. G. Skuratov ◽  
A. N. Lyzikov ◽  
E. V. Voropayev ◽  
O. V. Osipkina
Keyword(s):  
Liver Cirrhosis ◽  
Portal Hypertension ◽  
Genetic Analysis ◽  
Restriction Analysis ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Pathological Process ◽  
Interleukin 4 ◽  
Blood Leukocytes ◽  
Tnf Α

Objective: to assess the association of polymorphism of the markers of interleukin-4 (IL-4) and tumor necrosis factor (TNF-α) in patients with liver cirrhosis with severe pathological process. Material and methods. The research material was DNA isolated from blood leukocytes of the patients. To identify SNP data, the PCR method was chosen with subsequent restriction analysis and electrophoretic detection by horizontal gel electrophoresis and amplification with visualization of the obtained results. Results. The performed molecular and genetic analysis of IL-4 and TNF-α gene polymorphisms made it possible to assume a relation between the frequency of polymorphic genotypes and alleles of the investigated genes and the severity of liver cirrhosis and portal hypertension, which can be used to diagnose and predict the severe course of the disease.

scholarly journals Monitoring of rabies in wild animals in the Kirov region after oral immunization

2016 ◽  
Vol 61 (4) ◽  
pp. 186-192 ◽  
Author(s):  
O. N. Zaykova ◽  
T. V. Grebennikova ◽  
A. L. Elakov ◽  
K. S. Kochergin-Nikitsky ◽  
T. I. Aliper ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Vaccine Strain ◽  
Oral Vaccine ◽  
Molecular Genetic ◽  
Genetic Research ◽  
Oral Immunization ◽  
Molecular Genetic Analysis ◽  
Wild Isolate ◽  
Field Isolates ◽  
Pcr Techniques

This work presents the results of the molecular genetic research on genomes of field isolates of the rabies virus circulating in the territory of the Kirov region in order to analyze the phylogenetic relationship between the wild isolate genomes and to determine the possible reversion of the vaccine strain of the rabies virus used in the oral vaccine to virulent variant. We studied 24 brain samples from wild carnivores shot after oral immunization of the area with Rabivak-O/333. A bait with the vaccine provided by the Veterinary Service of the Kirov was also studied. All samples were found to be positive for the presence of the rabies virus as established by FAT and RT-PCR techniques. Phylogenetic analysis of N genome fragments of the rabies virus showed that the field isolates from the Kirov regions were genetically close to the field isolates from Buryatia 2012. Analysis of G genome fragments showed that the Kirov field isolates were close to the isolates from Lipetsk (2011), as well as to the Ukrainian isolates (2006 and 2010). Molecular genetic analysis of the gene fragments N and G for the field isolates and fragments of the genome of the rabies virus vaccine did not reveal any reversion to the virulent vaccine strain.

scholarly journals Risk factors of thrombotic complications in patients with single functional ventricle

2019 ◽  
Vol 64 (2) ◽  
pp. 68-74
Author(s):  
Yu. G. Lugacheva ◽  
I. V. Kulagina ◽  
I. A. Kovalev ◽  
Ye. V. Krivoschekov ◽  
O. S. Yanulevich ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Factor Vii ◽  
Molecular Genetic Testing ◽  
Heterozygous Genotype ◽  
Molecular Genetic Study ◽  
Hemostatic System ◽  
Factor Ii ◽  
Thrombotic Complications ◽  
Polymorphic Variants

Objective: to analyze the parameters of the hemostasis system and the results of molecular genetic testing in patients with a single functional ventricle. The study included 102 patients. All the patients underwent a staged surgical hemodynamic correction of a single functional ventricle. The authors performed a retrospective analysis of patient records in order to identify the episodes of thrombosis. The incidence of thrombotic complications at different stages of hemodynamic correction in the examined patients with a single functional ventricle was 12.7%. The indicators of plasma link hemostasis in the observed patients have been characterized by a balance of hemostatic reactions in the group of children with thrombosis and without. The results of a molecular genetic study demonstrated that the carrier of the heterozygous genotype of 20210GA factor II gene in patients with a single functional ventricle increased the risk of thrombotic complications 16 times (15.4% in patients with thrombosis versus 1.1% in the group without thrombosis; odds ratio 16.0; 95% confidence interval 1.34–191.24; p=0.028). All patients with thrombosis in the history revealed a homozygous condition according to variant 10976GG factor VII gene (p=0.017). Conclusion: molecular genetic analysis of polymorphic variants of the hemostatic system in patients with a single functional ventricle is required to predict the risk, timely prevention and correction of thrombotic complications during the surgical treatment of congenital heart disease.

scholarly journals Early lesion in the urinary system of a patient with DIDMOAD syndrome

2013 ◽  
Vol 59 (1) ◽  
pp. 18-22
Author(s):  
D P Grishina ◽  
L I Zil'berman ◽  
E Iu Zakharova ◽  
P G Tsygankova ◽  
I É Volkov ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Urinary System ◽  
Homozygous State ◽  
Molecular Genetic Study ◽  
Didmoad Syndrome ◽  
Early Lesion ◽  
Wfs1 Gene ◽  
First Time ◽  
Further Development

The authors present a case report of DIDMOAD syndrome in a girl accompanied by the early development of a severe lesion in the urinary system. The molecular-genetic analysis of the Wfs1 gene revealed thhec.1009A>C,p.T337P missense mutation in exon 8 in the homozygous state. The brother of the patient developed diabetes mellitus that for the first time manifested itself at the age of 6 years. The molecular-genetic study of the boy undertaken bearing in mind his genetically aggravated anamnesis revealed the analogous mutation; this finding maybe used to predict further development of the disease.

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