Diagnosis of Lyme Borreliosis With a Novel, Seminested Real-Time Polymerase Chain Reaction Targeting the 5S-23S Intergenic Spacer Region

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Adna Podbićanin-Ziburt ◽  
Thomas M. Falk ◽  
Dieter Metze ◽  
Almut Böer-Auer
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Lyme Borreliosis ◽  
Intergenic Spacer ◽  
Chain Reaction ◽  
Intergenic Spacer Region ◽  
Polymerase Chain

scholarly journals Differentiation of Moraxella Bovoculi sp. nov. from other Coccoid Moraxellae by the Use of Polymerase Chain Reaction and Restriction Endonuclease Analysis of Amplified DNA

2007 ◽  
Vol 19 (5) ◽  
pp. 532-534 ◽  
Author(s):  
John A. Angelos ◽  
Louise M. Ball
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Restriction Analysis ◽  
Intergenic Spacer ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Gram Negative ◽  
Intergenic Spacer Region ◽  
Polymerase Chain ◽  
Moraxella Bovoculi

Moraxella oris was historically the only coccoid Moraxella identified in cultures of ocular fluid from cattle with infectious bovine keratoconjunctivitis (IBK) and could be morphologically and biochemically differentiated from Moraxella bovis. Moraxella bovoculi sp. nov. is a recently characterized Moraxella isolated from ulcerated eyes of calves with IBK in northern California in 2002. Like Moraxella ovis, M. bovoculi sp. nov. is a gram-negative coccus/diplococcus. All 18 original isolates of M. bovoculi sp. nov. possessed phenylalanine deaminase (PADase) activity and could therefore be differentiated from M. ovis and M. bovis. During the characterization of 44 additional isolates of hemolytic gram-negative cocci that were cultured from ulcerated eyes of IBK-affected calves, 2 PADase-negative isolates were identified that could not be differentiated biochemically from M. ovis; however, the DNA sequence of the 16S-23S intergenic spacer region (ISR) of the isolates matched the 16S-23S ISR DNA sequence of M. bovoculi sp. nov. To facilitate the identification of PADase-negative moraxellae, a polymerase chain reaction (PCR) coupled with restriction enzyme digestion analysis of amplified DNA was developed. Amplification of the 16S-23S ISR followed by AfaI digestion of amplified DNA could differentiate M. bovoculi sp. nov. from M. ovis and other moraxellae. The DNA sequence analysis of the amplified 16S-23S ISR from the 42 PADase-positive isolates of hemolytic gram-negative cocci indicated that all were M. bovoculi sp. nov. and all possessed an AfaI site. A PCR coupled with restriction analysis of amplified DNA can aid in identifying M. bovoculi sp. nov.

Rapid Polymerase Chain Reaction/DNA Probe Membrane-Based Assay for the Detection of Listeria and Listeria monocytogenes in Food

2000 ◽  
Vol 63 (3) ◽  
pp. 337-342 ◽  
Author(s):  
L. O'CONNOR ◽  
J. JOY ◽  
M. KANE ◽  
T. SMITH ◽  
M. MAHER
Keyword(s):  
Polymerase Chain Reaction ◽  
Microbiological Quality ◽  
Intergenic Spacer ◽  
Pcr Primers ◽  
Dna Probe ◽  
23S Rrna ◽  
Chain Reaction ◽  
Intergenic Spacer Region ◽  
Rapid Polymerase Chain Reaction ◽  
Polymerase Chain

We describe the development of polymerase chain reaction (PCR)/DNA probe membrane-based colorimetric assays for the detection and identification of Listeria and L. monocytogenes. PCR primers designed from the 16S to 23S rRNA intergenic spacer region amplified products that were reverse hybridized to membrane-bound oligonucleotide probes specific for Listeria and L. monocytogenes with a detection limit of 1 to 10 CFU/25 ml in inoculated raw and pasteurized milk samples. These qualitative assays have the potential to be integrated into testing laboratories for monitoring the microbiological quality of foods.

Genus- and species-specific detection ofListeria monocytogenesusing polymerase chain reaction assays targeting the 16S/23S intergenic spacer region of the rRNA operon

1996 ◽  
Vol 42 (11) ◽  
pp. 1155-1162 ◽  
Author(s):  
Tom Graham ◽  
Elizabeth J. Golsteyn-Thomas ◽  
Victor P. J. Gannon ◽  
James E. Thomas
Keyword(s):  
Polymerase Chain Reaction ◽  
Listeria Monocytogenes ◽  
Sequence Data ◽  
Intergenic Spacer ◽  
23S Rrna ◽  
Chain Reaction ◽  
Nucleotide Sequence Data ◽  
Intergenic Spacer Region ◽  
Polymerase Chain ◽  
Species Specific

In this study, the 16S/23S rRNA intergenic spacer (IGS) regions of six Listeria species were examined. DNA bands of 590 and 340 bp were observed following polymerase chain reaction (PCR) amplification of DNA from Listeria monocytogenes, Listeria innocua, Listeria seeligeri, Listeria welshimeri, and Listeria ivanovii strains with generic rRNA IGS oligonucleotide primers. For strains of Listeria grayi subspp. grayi and murrayi, DNA band sizes of 550 and 340 bp were observed with this primer pair. DNA bands of these sizes were not observed for other Gram-negative or -positive bacteria in this PCR assay. Four RsaI digestion profiles were noted for the Listeria PCR products. Listeria monocytogenes strains had one profile; L. innocua strains had a second; L. seeligeri, L. welshimeri, and L. ivanovii strains had a third; and L. grayi strains had a fourth. The small and large 16S/23S rRNA IGSs of L. monocytogenes ATCC 15313 were identical in the first 58 5′ and the last 169 3′ nucleotides. However, the large rRNA IGS contained a central 267-bp region with tRNAIleand tRNAAlagenes. Large rRNA 16S/23S IGS nucleotide sequence data has not been previously reported. This data was used to develop novel Listeria genus-specific and L. monocytogenes species-specific PCR assays.Key words: polymerase chain reaction, 16S/23S rRNA intergenic spacer region, Listeria monocytogenes.

scholarly journals Determination of Locust Bean Gum and Guar Gum by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism Analysis

2001 ◽  
Vol 84 (1) ◽  
pp. 89-100 ◽  
Author(s):  
Kerstin Meyer ◽  
Christelle Rosa ◽  
Claudia Hischenhuber ◽  
Rolf Meyer
Keyword(s):  
Polymerase Chain Reaction ◽  
Guar Gum ◽  
Intergenic Spacer ◽  
Ice Cream ◽  
Locust Bean Gum ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Intergenic Spacer Region ◽  
Polymerase Chain ◽  
Locust Bean

Abstract A polymerase chain reaction (PCR) was developed to differentiate the thickening agents locust bean gum (LBG) and the cheaper guar gum in finished food products. Universal primers for amplification of the intergenic spacer region between trnL 3’ (UAA) exon and trnF (GAA) gene in the chloroplast (cp) genome and subsequent restriction analysis were applied to differentiate guar gum and LBG. The presence of <5% (w/w) guar gum powder added to LBG powder was detectable. Based on data obtained from sequencing this intergenic spacer region, a second PCR method for the specific detection of guar gum DNA was also developed. This assay detected guar gum powder in LBG in amounts as low as 1% (w/w). Both methods successfully detected guar gum and/or LBG in ice cream stabilizers and in foodstuffs, such as dairy products, ice cream, dry seasoning mixes, a finished roasting sauce, and a fruit jelly product, but not in products with highly degraded DNA, such as tomato ketchup and sterilized chocolate cream. Both methods detected guar gum and LBG in ice cream and fresh cheese at levels <0.1%.

scholarly journals Ribosomal Intergenic Spacer: A Polymerase Chain Reaction Diagnostic for Meloidogyne chitwoodi, M. fallax, and M. hapla

2002 ◽  
Vol 92 (8) ◽  
pp. 884-892 ◽  
Author(s):  
J. Wishart ◽  
M. S. Phillips ◽  
V. C. Blok
Keyword(s):  
Polymerase Chain Reaction ◽  
Length Polymorphism ◽  
Polymerase Chain Reaction Amplification ◽  
Intergenic Spacer ◽  
Chain Reaction ◽  
Meloidogyne Chitwoodi ◽  
Intergenic Spacer Region ◽  
Meloidogyne Spp ◽  
Reaction Amplification ◽  
Polymerase Chain

Polymerase chain reaction amplification of the intergenic spacer region between the 5S and 18S genes from Meloidogyne chitwoodi, M. fallax, and M. hapla enabled these three important temperate species to be differentiated. Length polymorphism was found between M. chitwoodi and M. fallax as a result of differing numbers of short repeats located between the 5S and 18S genes. The presence of the 5S gene within the rDNA cistrons was confirmed in the Meloidogyne spp. included in this study. The region between the 28S and 5S genes for M. chitwoodi and M. fallax was short and lacked variability in repeated sequences compared with the main tropical Meloidogyne spp. and M. hapla. Differences in the number of these repeats resulted in intraspecific length polymorphism for M.hapla

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