Rapid Detection and Identification of Lactobacillus spp. in Dairy Products by Using the Polymerase Chain Reaction

1996 ◽  
Vol 59 (10) ◽  
pp. 1031-1036 ◽  
Author(s):  
MARYANNE DRAKE ◽  
CHRISTOPHER L. SMALL ◽  
KEMET D. SPENCE ◽  
BARRY G. SWANSON
Keyword(s):  
Polymerase Chain Reaction ◽  
Dairy Products ◽  
Lactobacillus Helveticus ◽  
Lactobacillus Delbrueckii ◽  
Lactobacillus Species ◽  
Chain Reaction ◽  
Bacterial Dna ◽  
Detection And Identification ◽  
Polymerase Chain ◽  
Species Specific

Species-specific primers for use in the polymerase chain reaction (PCR) were designed to differentially amplify DNA from the common dairy lactobacillus species Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus helveticus, and Lactobacillus acidophilus. A method for rapid extraction of bacterial DNA from dairy products was developed. The sensitivity of bacterial DNA extraction from food and subsequent amplification by PCR was 100 cells total. Lactobacillus DNA was extracted and identified from commercial yoghurts, acidophilus milk, and cheeses. The methodology allows the presumptive identification of dairy lactobacilli in less than 6 hours.

Identification and determination of relatedness of lactobacilli using different DNA amplification methods

2012 ◽  
Vol 66 (9) ◽  
Author(s):  
Kristýna Turková ◽  
Bohuslav Rittich ◽  
Alena Španová
Keyword(s):  
Polymerase Chain Reaction ◽  
Dairy Products ◽  
Lactobacillus Rhamnosus ◽  
Dna Amplification ◽  
Term Infant ◽  
Lactobacillus Helveticus ◽  
Chain Reaction ◽  
Chain Reactions ◽  
Polymerase Chain ◽  
Species Specific

AbstractSeveral DNA amplification-based methods were used for identification and evaluation of the relation between lactobacilli isolated from breastfed full-term infant faeces (31 strains), dairy products (5 strains) and silage (1 strain). Twenty-seven strains isolated from infant faeces were identified as Lactobacillus rhamnosus (9), Lactobacillus gasseri (6), Lactobacillus paracasei (4), Lactobacillus fermentum (4), Lactobacillus salivarius (2), Lactobacillus plantarum (1), and Lactobacillus helveticus (1) using 10 species-specific polymerase chain reactions (PCRs), multiplex PCR for the Lactobacillus casei group, and sequencing of 16S rDNA. Four strains were not identified. Six strains isolated from dairy products and silage were identified as Lactobacillus rhamnosus. A repetitive extragenic palindromic polymerase chain reaction (rep-PCR) with primer (GTG)5 and a randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) with primer M13 were used for confirmation of species identification. Fingerprints were used for evaluation of the relatedness of lactobacilli. Differences between strains from infant faeces, dairy products, and silage were not detected.

Identification of starter cultures in thermally treated plain yogurt using gene probes and polymerase chain reaction

1996 ◽  
Vol 63 (4) ◽  
pp. 607-613 ◽  
Author(s):  
Sonja Lick ◽  
Martina Keller ◽  
Uli Krusch ◽  
Knut J. Heller
Keyword(s):  
Polymerase Chain Reaction ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Lactobacillus Delbrueckii ◽  
Agarose Gel Electrophoresis ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Specific Hybridization ◽  
Polymerase Chain ◽  
Species Specific

SummaryPlain yogurts (set type) fermented by three different starter cultures were subjected to different heat treatments (20 s at 72, 80 or 100 ·C, or 30 min at 60, 70, 80 or 100 ·C). DNA was extracted from each yogurt and analysed by agarose gel electrophoresis, by species-specific hybridization and by primer-specific polymerase chain reaction (PCR). Obvious degradation of DNA could be observed after heating at 80 ·C for 30 min and at 100 ·C for 20 s and 30 min. Identification ofLactobacillus delbrueckiiusing a species-specific hybridization fragment could be carried out in yogurt treated at 100 ·C for 20 s or for 30 min at lower temperatures. After treatment for 30 min at 100 ·C identification by hybridization was no longer possible. However, under the same conditions identification of starter microorganisms was still possible by primer-specific PCR, and this was demonstrated as an example forStreptococcus thermophilus.

scholarly journals Detection and Identification of Four Common Rust Pathogens of Cereals and Grasses Using Real-Time Polymerase Chain Reaction

2007 ◽  
Vol 97 (6) ◽  
pp. 717-727 ◽  
Author(s):  
C. W. Barnes ◽  
L. J. Szabo
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Internal Control ◽  
Chain Reaction ◽  
Detection And Identification ◽  
Pasture Grasses ◽  
Significant Yield ◽  
Polymerase Chain ◽  
Species Specific ◽  
Rust Pathogens

Puccinia spp. are widespread pathogens of cereals and grasses that annually cause significant yield losses worldwide, especially in barley, oat, and wheat. Urediniospore morphology and early symptom development have limited usefulness for distinguishing Puccinia spp. Therefore, we developed real-time polymerase chain reaction assays for rapid detection of the four rust pathogen species, Puccinia graminis (Pers.:Pers.), P. striiformis (Westend.), P. triticina (Eriks.), and P. recondita (Roberge ex Desmaz.). Duplex assays were constructed for the nuclear rDNA gene, using the variable internal transcribed spacer 1 (ITS1) region to distinguish between species, and the conserved 28S region as an internal control. Species-specific ITS1 primer/probe sets were highly specific and could detect <1 pg of DNA. The species-specific primer/probe sets showed positive results over a linear range of DNA five orders of magnitude or greater. Specificity of the assays was tested using multiple collections representing a range of races and formae speciales within a species. Additionally, assay specificity was evaluated by testing a range of other grass rust pathogens, as well as other fungi. The 28S primer/probe combination was successful in detecting all Puccinia spp. tested within the duplex assays, validating the integrity of each assay. Finally, the assays were used to identify unknown rust fungi infecting pasture grasses.

Routine identification of four sympatric species of calanoid copepods Pseudocalanus spp. in the Atlantic Arctic using a species-specific polymerase chain reaction

2020 ◽  
Vol 48 (1) ◽  
pp. 62-72
Author(s):  
E. A. Ershova
Keyword(s):  
Polymerase Chain Reaction ◽  
Life Cycles ◽  
The Arctic ◽  
Relative Distribution ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Routine Identification ◽  
Polymerase Chain ◽  
Species Specific ◽  
Plankton Communities

Сalanoid copepods of the genus Pseudocalanus play an important role in the plankton communities of the Arctic and boreal seas, often dominating in numbers and constituting a significant proportion of the biomass of zooplankton. Despite their high presence and significance in the shelf plankton communities, species-specific studies of the biology of these are significantly hampered by extremely small morphological differences between them, especially at the juvenile stages, at which they are virtually indistinguishable. In this paper, we describe a new, routine and low-cost molecular method for identifying all Pseudocalanus species found in the Atlantic sector of the Arctic: the Arctic P. acuspes, P. minutus and the boreal P. moultoni and P. elongatus, and apply it to describe the relative distribution of these species in four locations of the Arctic and sub-Arctic. With this method, species-specific polymerase chain reaction (ssPCR), mass identification of individuals of any developmental stage, including nauplii, is possible. This method can serve as an excellent tool for studying the species-specific biology of this group, describing their life cycles, as well as monitoring changes in Arctic marine ecosystems under the influence of changing climate.

scholarly journals A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Dna Markers ◽  
5S Rdna ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Species Specific ◽  
Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

Detection and identification of Candida spp. in human serum by LightCycler® real-time polymerase chain reaction

2008 ◽  
Vol 60 (3) ◽  
pp. 263-271 ◽  
Author(s):  
Catherine Dunyach ◽  
Sébastien Bertout ◽  
Cécile Phelipeau ◽  
Pascal Drakulovski ◽  
Jacques Reynes ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Serum ◽  
Real Time ◽  
Chain Reaction ◽  
Candida Spp ◽  
Detection And Identification ◽  
Polymerase Chain

Development of a Multiplex Polymerase Chain Reaction Assay for Detection and Differentiation of Staphylococcus aureus in Dairy Products†

2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Dairy Products ◽  
Multiplex Polymerase Chain Reaction ◽  
Skim Milk ◽  
Cheddar Cheese ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.

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