scholarly journals Genotyping of the Pseudorabies Virus by Multiplex PCR Followed by Restriction Enzyme Analysis

2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Antônio Augusto Fonseca ◽  
Cristina Gonçalves Magalhães ◽  
Érica Bravo Sales ◽  
Régia Maria D'Ambros ◽  
Janice Ciacci-Zanella ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Restriction Enzyme ◽  
Pseudorabies Virus ◽  
Virus Isolation ◽  
Restriction Analysis ◽  
Infectious Agent ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Restriction Sites ◽  
Herpesvirus 1

Suid herpesvirus 1 (SuHV-1) is the causative agent of Aujeszky's disease. The infectious agent has only one serotype, but it was classified by restriction enzyme analysis of the whole genome into four genotypes, named I to IV. The aim of this study was to standardize a rapid method for genotyping SuHV-1 without virus isolation, using a multiplex-PCR followed by enzymatic restriction analysis. The complete genome of the virus was analyzed in silico to determine the restriction sites for the enzyme BamHI. Primers were designed to flank sites with emphasis on certain points of differentiation of genotypes. The standard PCRs were able to detect the SuHV-1 and also to differentiate genotypes from brain tissue of infected pigs. The BamHI-PCR is a rapid, practical, and sensitive way to genotype SuHV-1.

scholarly journals A simple method for detection of mutations in amino acid 452 of the Spike protein of SARS-CoV-2 using restriction enzyme analysis

2021 ◽  
Vol 62 (4) ◽  
pp. 371-377
Author(s):  
Rossana C. Jaspe ◽  
Yoneira Sulbaran ◽  
Mariana Hidalgo ◽  
Mariana Hidalgo ◽  
Carmen L. Loureiro ◽  
...  
Keyword(s):  
Immune Response ◽  
Amino Acid ◽  
Restriction Enzyme ◽  
Restriction Analysis ◽  
Spike Protein ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Simple Method ◽  
Spike Gene ◽  
Detection Of Mutations

Variants of Concern or Interest of SARS-CoV-2 (VOC or VOI), the coronavirus responsible for COVID-19, have emerged in several countries. Mutations in the amino acid 452 of the Spike protein are particularly important and associated with some of these variants: L452R, present in Delta VOC, and L452Q, present in Lambda VOI. These mutations have been associated with both increased infectivity and evasion of protective immune response. A search on GISAID to detect the number of sequences harboring the L452R mutation and the frequency of Delta VOC among them, showed that since August 2021, most of these sequences belong to the Delta VOC. Restriction enzyme analysis is proposed as a rapid method to detect L452R. A small amplicon from the Spike gene was digested with MspI. A 100% concordance was observed between digestion and sequencing results. The mutation L452Q can also be detected by restriction analysis, allowing the identification of putative Lambda VOIs. The proposed methodology, which allows screening of a great number of samples, could provide a faster information on the prevalence of Delta VOC cases.

scholarly journals Specific Detection of Xanthomonas axonopodis pv. dieffenbachiae in Anthurium (Anthurium andreanum) Tissues by Nested PCR

2006 ◽  
Vol 72 (2) ◽  
pp. 1072-1078 ◽  
Author(s):  
Isabelle Robène-Soustrade ◽  
Philippe Laurent ◽  
Lionel Gagnevin ◽  
Emmanuel Jouen ◽  
Olivier Pruvost
Keyword(s):  
Diagnostic Tool ◽  
Nested Pcr ◽  
Restriction Analysis ◽  
Detection Threshold ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Amplification Product ◽  
Xanthomonas Axonopodis ◽  
Sequence Characterized Amplified Region ◽  
Pcr Test

ABSTRACT Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (103 CFU ml−1) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

Long interspersed repeated DNA (LINE) causes polymorphism at the rat insulin 1 locus

1985 ◽  
Vol 5 (9) ◽  
pp. 2197-2203
Author(s):  
M S Lakshmikumaran ◽  
E D'Ambrosio ◽  
L A Laimins ◽  
D T Lin ◽  
A V Furano
Keyword(s):  
Dna Sequence ◽  
Restriction Enzyme ◽  
Allelic Variation ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Repeated Dna ◽  
Coding Region ◽  
Sequence Determination ◽  
Polymorphic Region ◽  
Or Gene

The insulin 1, but not the insulin 2, locus is polymorphic (i.e., exhibits allelic variation) in rats. Restriction enzyme analysis and hybridization studies showed that the polymorphic region is 2.2 kilobases upstream of the insulin 1 coding region and is due to the presence or absence of an approximately 2.7-kilobase repeated DNA element. DNA sequence determination showed that this DNA element is a member of a long interspersed repeated DNA family (LINE) that is highly repeated (greater than 50,000 copies) and highly transcribed in the rat. Although the presence or absence of LINE sequences at the insulin 1 locus occurs in both the homozygous and heterozygous states, LINE-containing insulin 1 alleles are more prevalent in the rat population than are alleles without LINEs. Restriction enzyme analysis of the LINE-containing alleles indicated that at least two versions of the LINE sequence may be present at the insulin 1 locus in different rats. Either repeated transposition of LINE sequences or gene conversion between the resident insulin 1 LINE and other sequences in the genome are possible explanations for this.

Restriction Enzyme Analysis (REA) of Group A Streptococcal (GAS) M-Serotypes 1, 3, and 28

1997 ◽  
pp. 221-223 ◽  
Author(s):  
Dwight R. Johnson ◽  
Cheryl L. Romana ◽  
Carey D. Rehder ◽  
Joanne Dehnbostel ◽  
Edward L. Kaplan
Keyword(s):  
Restriction Enzyme ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Group A

scholarly journals Identificação de micobactérias não tuberculosas isoladas de sítios estéreis em pacientes em um hospital universitário na cidade do Rio de Janeiro

2011 ◽  
Vol 37 (4) ◽  
pp. 521-526 ◽  
Author(s):  
Simone Gonçalves Senna ◽  
Ana Grazia Marsico ◽  
Gisele Betzler de Oliveira Vieira ◽  
Luciana Fonseca Sobral ◽  
Philip Noel Suffys ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Restriction Enzyme ◽  
Rio De Janeiro ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis

OBJETIVO: Identificar micobactérias não tuberculosas (MNT) isoladas de sítios estéreis em pacientes internados no Hospital Universitário Clementino Fraga Filho, Rio de Janeiro (RJ) entre 2001 e 2006. MÉTODOS: Durante o período do estudo, 34 isolados de MNT de sítios estéreis de 14 pacientes, a maioria HIV positivos, foram submetidos a identificação fenotípica e hsp65 PCR-restriction enzyme analysis (PRA, análise por enzimas de restrição por PCR do gene hsp65). RESULTADOS: A maioria dos isolados foi identificada como Mycobacterium avium, seguida por M. monacense, M. kansasii e M. abscessus em menores proporções. CONCLUSÕES: A combinação de PRA, um método relativamente simples e de baixo custo, com algumas características fenotípicas pode fornecer a identificação correta de MNT na rotina de laboratórios clínicos.

RESTRICTION ENZYME ANALYSIS OF HERPESVIRUS-2 DNA

The Lancet ◽  
1981 ◽  
Vol 318 (8260-8261) ◽  
pp. 1424 ◽  
Author(s):  
IsabelW Smith ◽  
N.J Maitland ◽  
J.F Peutherer ◽  
D.H.H Robertson
Keyword(s):  
Restriction Enzyme ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis
Sign in / Sign up

Export Citation Format

Share Document