scholarly journals Application of real-time PCR for DNA identification of ruminants in raw animal products

2018 ◽  
Vol 74 (1) ◽  
pp. 6061-2018
Author(s):  
ANNA WEINER ◽  
ILONA PAPROCKA ◽  
AGATA GOŁĘBIOWSKA ◽  
KRZYSZTOF KWIATEK
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Real Time Pcr ◽  
Animal Origin ◽  
Dna Identification ◽  
Animal Products ◽  
Modified Method ◽  
Pcr Method ◽  
Processed Products ◽  
By Products

According with current regulations it is possible export of processed animal proteins produced from tissues other animals than ruminants. In order to ensure appropriate control studies were undertaken to adopt of method real-time PCR used for analysis of processed products of animal origin. The material consisted of raw animal by-products of beef, pork and poultry. In the assays mitochondrial DNA was used. Modified method allows to detect the DNA of ruminants at the level of 0.025%. Based on obtained results of the validation studies were found that the real-time PCR method can be used in routine tests for identification of ruminant DNA in raw animal by-products..

Human Remains detection after Suicide Bombing by STR Analysis

1970 ◽  
Vol 15 (1) ◽  
pp. 53-57
Author(s):  
Majeed Arsheed Sabbah ◽  
Halah Khalid Ibrahim ◽  
Dhuha Salim Namaa ◽  
Sura Nabeel
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Human Remains ◽  
Animal Origin ◽  
Suicide Bombing ◽  
Dna Identification ◽  
Str Analysis ◽  
Excessive Heat ◽  
South Of Iraq ◽  
Genetic Analyzer

        Following suicide bombing, human remains usually collected for DNA identification. Human remains affected due to exposure to excessive heat and bomb solid, sharp pieces. The aim of this work is to study the possibility of DNA analyzing of human remains after suicide bombing. Fifteen human remains were received from a place of suicide bombing in Najaf City south of Iraq. The remains were severely affected and cannot recognize morphologically or as it is human or animal origin. DNA extracted from remains by organic method, then quantified by real time PCR kit then analyzed by Powerplex 21 kit using 3130XL Genetic Analyzer. The results showed that seven remains where analyzed successfully while the other remains failed for analysis for both real time PCR quantification or STR analysis. Four remains belong to two persons.  This study showed that suicide bombing affects negatively the most remains for STR analysis.

scholarly journals Methods of Detecting Meat Species in Food of Animal Origin

2018 ◽  
Vol 75 (2) ◽  
pp. 125
Author(s):  
Lavinia-Maria CHIŞ ◽  
Dan-Cristian VODNAR
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Product ◽  
Meat Products ◽  
Ease Of Use ◽  
Animal Origin ◽  
Processed Meat ◽  
Elisa Method ◽  
Food Of Animal Origin ◽  
Pcr Method

An important factor in the detection of falsification is the control of the composition of the meat at each stage of manufacturing the product. The PCR method is based on the study of proteins and meat nucleic acids used in food for the detection of animal species. Another technique is the Elisa method that works on the principle of identification and measurement of the quantity of molecules in a sample. There are several types of Elisa to increase specificity due to differences in structure and sample characteristics. By comparing the two methods used to identify the processed meat product species, Real Time PCR had the highest prediction as results. However, the Elisa method is more time efficient and easier to use. Real Time PCR is effective in identifying processed meat products that require low detection. The Elisa Kit is useful because of the ease of use.

A real-time PCR method for quantifying mixed cashmere and wool based on hair mitochondrial DNA

2014 ◽  
Vol 84 (15) ◽  
pp. 1612-1621 ◽  
Author(s):  
Minfeng Tang ◽  
Weiping Zhang ◽  
Hui Zhou ◽  
Jing Fei ◽  
Juan Yang ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Method

scholarly journals Detection of food hazards in foods: comparison of real time polymerase chain reaction and cultural methods

2016 ◽  
Vol 5 (1) ◽  
Author(s):  
Paolo Bonilauri ◽  
Lia Bardasi ◽  
Roberto Leonelli ◽  
Mattia Ramini ◽  
Andrea Luppi ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Reference Method ◽  
Food Samples ◽  
Alternative Methods ◽  
Animal Origin ◽  
Screening Methods ◽  
Molecular Screening ◽  
Cultural Methods ◽  
Pcr Method

Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time PCR are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisation. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyze the data collected, from 2012 to 2014, by Emilia Romagna Region in the field of <em>Piano Regionale Alimenti</em> (Food Regional Plan), during official controls monitoring food samples, of animal and other than animal origin. Records performed by combined methods of molecular screening of <em>Salmonella</em> spp., <em>Listeria monocytogenes</em> and thermophilic <em>Campylobacter</em> and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10.604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected <em>Salmonella</em>, <em>L. monocytogenes</em>, and thermophilic <em>Campylobacter</em> in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57 % of samples, respectively. In spite of the use of the same enrichment broth, the real time PCR method disclosed a percentage of positive samples that were negative to cultural examination ranging between 20 and 43%, with a PCR/culture ratio between 2.37 to 5.00. In conclusion, the results of this study pose a doubt about the sensitivity of the official cultural methods regarding the isolation of the three investigated foodborne pathogens. Moreover this study may be a useful tool for Veterinary Authorities to assess appropriate sampling plans to control the risk relating to the consumption of contaminated foods.

HISTOPHILUS SOMNI DNA IDENTIFICATION IN SEMEN OF BULLS BY REAL-TIME PCR METHOD

2020 ◽  
Vol 1 ◽  
pp. 58-60
Author(s):  
S.P. Yatsentyuk ◽  
◽  
D.A. Rudnyaev ◽  
Yu.I. Pobolelova ◽  
A.D. Kozlova ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Dna Identification ◽  
Histophilus Somni ◽  
Pcr Method

Development of a real-time PCR method for the simultaneous detection of soya and lupin mitochondrial DNA as markers for the presence of allergens in processed food

2011 ◽  
Vol 127 (2) ◽  
pp. 834-841 ◽  
Author(s):  
Antonio M. Gomez Galan ◽  
Marcel Brohée ◽  
Eugénia de Andrade Silva ◽  
Arjon J. van Hengel ◽  
Hubert Chassaigne
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Processed Food ◽  
Pcr Method

A multiplex real-time PCR method to detect and quantify mitochondrial DNA deletions in individual cells

2007 ◽  
Vol 370 (1) ◽  
pp. 127-129 ◽  
Author(s):  
Kim J. Krishnan ◽  
Andreas Bender ◽  
Robert W. Taylor ◽  
Douglass M. Turnbull
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Real Time Pcr ◽  
Dna Deletions ◽  
Pcr Method

Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

2020 ◽  
Vol 18 ◽  
Author(s):  
Pegah Shakib ◽  
Mohammad Reza Zolfaghari
Keyword(s):  
Streptococcus Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
False Negative ◽  
Cross Sectional Study ◽  
Laboratory Culture ◽  
Cross Sectional ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

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.

Development and application of a novel reverse transcription real-time PCR method for miR-499 quantification

2013 ◽  
Vol 46 (15) ◽  
pp. 1566-1571 ◽  
Author(s):  
Weidong Zheng ◽  
Yuwei Di ◽  
Yinghong Liu ◽  
Ge Huang ◽  
Youwei Zheng ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Pcr Method
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