scholarly journals Molecular Detection of Clostridium botulinum Type E Neurotoxin Gene in Smoked Fish by Polymerase Chain Reaction and Capillary Electrophoresis

1996 ◽  
Vol 79 (4) ◽  
pp. 861-865 ◽  
Author(s):  
Carl J Sciacchttano ◽  
Irvin N Hlrshfield
Keyword(s):  
Polymerase Chain Reaction ◽  
Capillary Electrophoresis ◽  
Dna Sequences ◽  
Clostridium Botulinum ◽  
Polymer System ◽  
Chain Reaction ◽  
Smoked Fish ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E ◽  
Polymerase Chain

Abstract The polymerase chain reaction (PCR), a rapid, sensitive technique for amplifying target DNA sequences of pathogenic microorgansims, was used to amplify Clostridium botulinum type E neurotoxin gene fragments in smoked fish. Other botulinal neurotoxin-producing strains, nontoxigenic strains, and food-related microorganisms did not yield nonspecificamplification products with this PCR assay. PCR products were analyzed by capillary electrophoresis (CE) using a low-viscosity entangled polymer system. Resolution, sensitivity, and DNA sizing accuracy were improved, and analytical times were markedly shortened. The PCR/CE assay detected the C. botulinum type E neurotoxin gene in as few as 10 cells. The technique to other foods may also be a valuable tool for detecting foodborne pathogens.

Detection of Clostridium botulinum type F using the polymerase chain reaction

1994 ◽  
Vol 8 (5) ◽  
pp. 365-373 ◽  
Author(s):  
Joseph L. Ferreira ◽  
Mostafa K. Hamdy ◽  
Steven G. McCay ◽  
Mark Hemphill ◽  
Nameer Kirma ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Botulinum ◽  
Chain Reaction ◽  
Botulinum Type ◽  
Polymerase Chain

scholarly journals Optimization of Polymerase Chain Reaction for Detection of Clostridium botulinum Type C and D in Bovine Samples

2007 ◽  
Vol 54 (8) ◽  
pp. 320-327 ◽  
Author(s):  
V. Prévot ◽  
F. Tweepenninckx ◽  
E. Van Nerom ◽  
A. Linden ◽  
J. Content ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Botulinum ◽  
Chain Reaction ◽  
Botulinum Type ◽  
Type C ◽  
Polymerase Chain

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

Polymerase chain reaction based mapping of rye involving repeated DNA sequences

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 621-626 ◽  
Author(s):  
Peter M. Rogowsky ◽  
Ken W. Shepherd ◽  
Peter Langridge
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Chromosome 1 ◽  
Repeated Dna ◽  
Chain Reaction ◽  
Pcr Marker ◽  
Banding Patterns ◽  
Repeated Dna Sequences ◽  
Cereal Rye ◽  
Polymerase Chain

A novel type of polymerase chain reaction (PCR) marker was developed for the mapping of cereal rye (Secale cereale). Primer pairs were synthesized targeting the insertion sites of three individual copies of the R173 family of rye specific repeated DNA sequences. While one primer was derived from a sequence within the respective R173 element, the second primer corresponded to a flanking region. The complex banding patterns obtained in rye allowed not only the mapping of the three R173 elements to certain chromosome regions of 1RS (the short arm of rye chromosome 1) but also the mapping of an additional 3–10 easily identifiable bands per primer pair to other rye chromosomes. Linkage mapping of a polymorphic 1R band derived from three rye cultivars demonstrated the presence of nonallelic, dominant markers in two independent crosses. Because of the high copy number of the R173 family (15 000 copies per diploid rye genome), its dispersion over the entire length of all chromosomes and the high number of markers obtained per primer pair, PCR markers based on the R173 family provide an almost unlimited source for well-spaced markers in rye mapping.Key words: polymerase chain reaction, mapping, repetitive DNA sequences, wheat, rye.

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