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[A molecular genetic analysis of spinal muscular atrophy (SMA) in families at high risk from different regions of Ukraine]

https://arctichealth.org/en/permalink/ahliterature33908
Source
Tsitol Genet. 1997 Nov-Dec;31(6):75-81
Publication Type
Article
Author
A Iu Ekshiian
L A Livshits
A M Bychkova
N A Afanas'eva
I R Bariliak
Source
Tsitol Genet. 1997 Nov-Dec;31(6):75-81
Language
Russian
Publication Type
Article
Keywords
Adult
Child
Chimera - genetics
DNA - genetics - isolation & purification
DNA Primers
Electrophoresis, Agar Gel
English Abstract
Exons - genetics
Gene Deletion
Heterozygote
Homozygote
Humans
Molecular Biology
Polymerase Chain Reaction - methods
Research Support, Non-U.S. Gov't
Risk factors
Spinal Muscular Atrophies of Childhood - genetics
Ukraine
Abstract
The results of DNA analysis of deletion in exons 7 and 8 of SMN gene, and exon 5 of NAIP gene in 24 SMA-families from Ukraine are presented. Deletions of SMN exons 7 and 8, or 7 were found in 46 (97.9%) of 47 SMA-chromosomes. A homozygous deletion of NAIP exon 5 was demonstrated in 4 (19%) of 21 SMA-families. The authors have demonstrated that in 2 SMA patients with homozygous deletion SMN exon 7 only, the remaining SMN exon 8 was a part of a chimeric CBCD41/SMN gene.
PubMed ID
9591348 View in PubMed
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Detection of Streptococcus pneumoniae DNA by using polymerase chain reaction and microwell hybridization with Europium-labelled probes.

https://arctichealth.org/en/permalink/ahliterature31551
Source
J Microbiol Methods. 2002 Aug;50(3):313-8
Publication Type
Article
Date
Aug-2002
Author
S. Rintamäki
A. Saukkoriipi
P. Salo
A. Takala
M. Leinonen
Author Affiliation
National Public Health Institute (KTL), P.O. Box 310, 90101, Oulu, Finland.
Source
J Microbiol Methods. 2002 Aug;50(3):313-8
Date
Aug-2002
Language
English
Publication Type
Article
Keywords
Bacteriological Techniques - methods
DNA, Bacterial - analysis
Electrophoresis, Agar Gel
Europium - chemistry
Evaluation Studies
Fluorescent Dyes - chemistry
Humans
Nucleic Acid Hybridization - methods
Polymerase Chain Reaction - methods
Sensitivity and specificity
Streptococcus pneumoniae - genetics - isolation & purification
Abstract
The present paper describes a novel modification of polymerase chain reaction (PCR) for the detection of Streptococcus pneumoniae DNA in clinical specimens. PCR was based on the detection of a 209-base pair segment of the S. pneumoniae pneumolysin gene. For the demonstration of the amplification product, microwell hybridization with a Europium-labelled oligonucleotide probe complementary to a biotinylated strand of the PCR product was performed, and the presence of the PCR product was monitored by time-resolved fluorescence (TRF) of the Europium chelate. The sensitivity of the assay for purified S. pneumoniae DNA was 50 fg DNA corresponding to 20 genome equivalents of S. pneumoniae DNA. The efficiency of the hybridization step was monitored by using known amounts of synthetic target oligonucleotides as standards. Sensitivity of 3 x 10(8) molecules per individual reaction well was achieved with a 30-min attachment time and a 3-h hybridization time.Detection of PCR-amplified products by the microwell hybridization technique and TRF was compared to agarose gel electrophoresis in 50 middle ear fluid samples obtained from children with acute otitis media. The agarose gel and TRF detection methods identified all culture-positive samples, but both were also positive for 55% of the culture-negative samples. The results suggest that the detection of amplified PCR products by microwell hybridization using Europium-labelled oligonucleotides is a reliable method for the demonstration of the pneumolysin gene fragment. Furthermore, the method is suitable for automation and, thus, for testing high numbers of samples. The clinical significance of the PCR findings remains to be studied.
PubMed ID
12031582 View in PubMed
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Sample preparation and DNA extraction procedures for polymerase chain reaction identification of Listeria monocytogenes in seafoods.

https://arctichealth.org/en/permalink/ahliterature11066
Source
Int J Food Microbiol. 1997 Apr 15;35(3):275-80
Publication Type
Article
Date
Apr-15-1997
Author
A. Agersborg
R. Dahl
I. Martinez
Author Affiliation
Norwegian Institute of Fisheries and Aquaculture N-9005 Tromso, Norway.
Source
Int J Food Microbiol. 1997 Apr 15;35(3):275-80
Date
Apr-15-1997
Language
English
Publication Type
Article
Keywords
Animals
Base Sequence
DNA, Bacterial - analysis - chemistry - genetics
Decapoda (Crustacea) - microbiology
Detergents - pharmacology
Electrophoresis, Agar Gel
Endopeptidase K - pharmacology
Fish Products - microbiology
Food Microbiology
Food Poisoning - diagnosis - epidemiology - etiology
Gene Amplification
Heat
Humans
Listeria monocytogenes - drug effects - genetics - isolation & purification
Muramidase - pharmacology
Norway - epidemiology
Octoxynol - pharmacology
Polymerase Chain Reaction - methods
Prevalence
Research Support, Non-U.S. Gov't
Sensitivity and specificity
Time Factors
Abstract
Five grams of seafood products were inoculated with one to 500 viable or 10(9) heat-killed cells of Listeria monocytogenes. The presence of the pathogen was detected by the polymerase chain reaction (PCR) with primers specific for fragments of the listeriolysin O (hly) gene (two sets) and for the invasion-associated protein (iap) gene (one set). For DNA preparation, boiling, either alone or in combination with lysozyme and proteinase K treatment, was not always sufficient to lyse L. monocytogenes, while treatment with Triton X-100 produced consistently good DNA suitable for amplification. To avoid false-negative and false-positive results, 48 h incubations were necessary and a subculturing step after an initial 24 h incubation greatly improved the results. The primers that amplified regions of the listeriolysin O gene gave clearer and stronger products than primers for the invasion-associated protein gene. Using this method we were able to detect one to five L. monocytogenes cells in 5 g of product in a total of 55 h.
PubMed ID
9105938 View in PubMed
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