scholarly journals DNA barcoding, species-specific PCR and real-time PCR techniques for the identification of six Tribolium pests of stored products

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Tao Zhang ◽  
Yi-Jiao Wang ◽  
Wei Guo ◽  
Dan Luo ◽  
Yi Wu ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Barcoding ◽  
Real Time Pcr ◽  
Stored Products ◽  
Specific Pcr ◽  
Species Specific ◽  
Pcr Techniques

Conventional and Real-Time PCR–Based Approaches for Molecular Detection and Quantitation of Bovine Species Material in Edible Gelatin

2005 ◽  
Vol 68 (11) ◽  
pp. 2420-2426 ◽  
Author(s):  
TAURAI TASARA ◽  
SANDRA SCHUMACHER ◽  
ROGER STEPHAN
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Molecular Detection ◽  
Raw Material ◽  
Spongiform Encephalopathy ◽  
Specific Pcr ◽  
Species Specific ◽  
Bovine Gelatin ◽  
Bovine Species ◽  
Pcr Primer

The majority of edible gelatin in Europe is derived from pigskin, but a significant portion is extracted from bovine tissue. Because of the bovine spongiform encephalopathy crisis, consumers might be concerned about the gelatin used in various products. To assure consumers of the quality and safety of edible gelatin, European Union directive 1999/724/EC described general guidelines for gelatin production, including requirements for documentary proof confirming that raw materials are from animals fit for human consumption. Analytical methods to confirm gelatin documentation or raw material animal species source in the finished product are lacking. In this study, several published species-specific PCR systems were evaluated as potential molecular methods for determining the origin of the raw material used in making gelatin. A recently validated bovine species-specific PCR primer set targeting the ATPase 8 subunit gene in bovine mitochondrial DNA was suitable for detection of bovine material in gelatin. This PCR primer set was optimized using conventional and real-time PCR approaches. An evaluation of these two PCR methods confirmed the high specificity for the adopted primer set in various gelatin matrices of known origin. The inclusion of bovine gelatin in pork or fish gelatin can be detected at 0.1 to 0.001%. These PCR assays are potential molecular detection tools that can be used to routinely detect bovine gelatin either alone or as an inclusion in gelatin made from other species.

scholarly journals SNP Typing Using Multiplex Real-Time PCR Assay for Species Identification of Forensically Important Blowflies and Fleshflies Collected in South Korea (Diptera: Calliphoridae and Sarcophagidae)

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Hari Jang ◽  
Sang Eon Shin ◽  
Kwang soo Ko ◽  
Seong Hwan Park
Keyword(s):  
South Korea ◽  
Real Time ◽  
Dna Barcoding ◽  
Real Time Pcr ◽  
Coi Gene ◽  
Nucleotide Sequencing ◽  
Morphological Identification ◽  
Growth Data ◽  
Important Species ◽  
Species Specific

Medicolegal entomology—a subfield of forensic entomology—is mainly used in medicolegal investigations to estimate the postmortem interval (PMI). The minimum PMI of a corpse invaded by necrophagous immature insects can be estimated because the PMI is near to or earlier than the oviposition time of the larvae that hatched and fed on the corpse. As the growth speeds of larvae differ depending on temperature and species, species-specific growth data are used to estimate the minimum PMI. While morphological identification of adult necrophagous flies can be done by a well-trained entomologist, identification of larvae is relatively difficult. Larvae can only be identified up to the family level and developmental stage by observing the posterior spiracles. For these reasons, the molecular biology method of DNA barcoding has been developed. DNA barcoding that targets the mitochondrial cytochrome c oxidase subunit I (COI) gene is commonly used. COI sequences are currently acquired using polymerase chain reaction (PCR) and Sanger sequencing, which are too time-consuming and complex for practical use in medicolegal investigations. To compensate for these limitations and facilitate the use of entomology for medicolegal investigation, we designed a multiplex real-time PCR system to identify nineteen forensically important species of Calliphoridae and Sarcophagidae flies collected in South Korea. In contrast to the Sanger nucleotide sequencing process, this technology only requires a one-step real-time PCR with melt curve analysis of amplicons generated by primers targeting species-specific single nucleotide polymorphisms (SNPs). Multiplex real-time PCR was performed for twelve species of Calliphoridae (four reactions) and for seven species of Sarcophagidae (three reactions). This assay is expected to make it easier and faster for investigating authorities to identify major species of necrophagous flies at beginning of investigation and to increase the utilization of entomological evidence in forensic investigations.

scholarly journals Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria

2020 ◽  
Vol 9 (1) ◽  
pp. 54
Author(s):  
Salem Belkessa ◽  
Daniel Thomas-Lopez ◽  
Karim Houali ◽  
Farida Ghalmi ◽  
Christen Rune Stensvold
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Intestinal Parasites ◽  
Giardia Duodenalis ◽  
Specific Pcr ◽  
Stool Samples ◽  
Pcr Targeting ◽  
Assemblage B

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.

scholarly journals Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Primers ◽  
Rrna Gene ◽  
Dekkera Bruxellensis ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Spoilage Yeast ◽  
Pcr Method ◽  
26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

Species-specific real-time PCR cell number quantification of the bloom-forming cyanobacterium Planktothrix agardhii

2012 ◽  
Vol 194 (9) ◽  
pp. 749-757 ◽  
Author(s):  
Catarina Churro ◽  
Paulo Pereira ◽  
Vitor Vasconcelos ◽  
Elisabete Valério
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Cell Number ◽  
Planktothrix Agardhii ◽  
Species Specific

Identification of L. casei and L. rhamnosus in the dairy products using Real-Time PCR assay

2015 ◽  
Vol 4 (5) ◽  
pp. 222-225
Author(s):  
K. G. Li ◽  
G. P. Pogossian ◽  
A. K. Moldagulova ◽  
E. E. Bekenova ◽  
A. Abdikadirova ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Dairy Products ◽  
Production Control ◽  
High Efficiency ◽  
High Specificity ◽  
Industrial Test ◽  
Fermented Dairy Products ◽  
Species Specific ◽  
Fermented Dairy

  Lactobacilli are essential and important biological objects used in food pro-duction and medicine. One of the sufficient problems is fast, reliable and highly specific identification of lactobacilli in the scientific research and cur-rent production control. We represent two species-specific real-time PCR in the present study to discriminate L. rhamnosus and L. casei basing on the unique peptidoglycan-hydrolase genes p40 and p75 respectively. PCR pri-mers and probes were designed to provide high specificity discrimination via high temperature of PCR annealing stage. High efficiency of the reactions is provided by the size of amplified DNA fragments minimization. Reliable re-producibility of the target sequences amplification and fluorescence detec-tion provide a basis for the future creation of industrial test-systems for op-erational control in the production of fermented dairy products.

Quantitative Detection of Species-Specific DNA in Feedstuffs and Fish Meals

2005 ◽  
Vol 68 (6) ◽  
pp. 1217-1221 ◽  
Author(s):  
PAVEL KRCMAR ◽  
EVA RENCOVA
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Dna Sequences ◽  
Real Time Pcr ◽  
Single Species ◽  
Absolute Quantification ◽  
Quantitative Detection ◽  
Vertebrate Species ◽  
Target Species ◽  
Species Specific

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived material in concentrate feed mixtures and fish meals.

scholarly journals Characterization of Plasmodium ovale curtisi and P. ovale wallikeri in Western Kenya Utilizing a Novel Species-specific Real-time PCR Assay

2015 ◽  
Vol 9 (1) ◽  
pp. e0003469 ◽  
Author(s):  
Robin H. Miller ◽  
Clifford O. Obuya ◽  
Elizabeth W. Wanja ◽  
Bernhards Ogutu ◽  
John Waitumbi ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Novel Species ◽  
Plasmodium Ovale ◽  
Pcr Assay ◽  
Western Kenya ◽  
Plasmodium Ovale Curtisi ◽  
Species Specific

scholarly journals Rapid Species Identification of Cooked Poisonous Mushrooms by Using Real-Time PCR

2008 ◽  
Vol 74 (10) ◽  
pp. 3306-3309 ◽  
Author(s):  
Kazuhiko Maeta ◽  
Tomoya Ochi ◽  
Keisuke Tokimoto ◽  
Norihiro Shimomura ◽  
Nitaro Maekawa ◽  
...  
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Specific Test ◽  
Specific Identification ◽  
Specific Fluorescence ◽  
Species Specific ◽  
Poisonous Mushrooms

ABSTRACT Species-specific identification of the major cooked and fresh poisonous mushrooms in Japan was performed using a real-time PCR system. Specific fluorescence signals were detected, and no nonspecific signals were detected. Therefore, we succeeded in developing a species-specific test for the identification of poisonous mushrooms within 1.5 h.

Sign in / Sign up

Export Citation Format

Share Document