scholarly journals Comparison of real time PCR and conventional PCR by identifying genomic DNA of bovine and porcine

2021 ◽  
Vol 23 (2) ◽  
pp. 63-71
Author(s):  
Muhammad Abdurrahman Munir ◽  
Ahlam Inayatullah
Keyword(s):  
Real Time ◽  
Genomic Dna ◽  
Dna Analysis ◽  
Meat Products ◽  
Poultry Meat ◽  
Cyt B ◽  
Conventional Pcr ◽  
Specific Difference ◽  
Bovine Tissue ◽  
Dna Primers

Bovine and porcine are poultry meat that consumed worldwide particularly in Southeast Asia.Both of them are prone to food counterfeit owing to several factors such as price, appetite and Halal status. Sensitive and selective analytical methods are required to control meat products that distributed to markets. This paper studied the sensitivity between real – time and conventional PCR or known as qPCR and cPCR, respectively. Bovine and porcine were samples used to verify the sensitivity of them. Nevertheless, those instruments did not show a specific difference during DNA analysis of bovine and porcine. In conventional PCR, two pairs of DNA primers targeted cytochrome b (Cyt b) was analyzed, resulting of 120 and 131 amplicons, respectively. While qPCR applied to analyze porcine and bovine DNA. The detection limit of qPCR after porcine and bovine analysis were at 0.004 and 0.007 µg/µL, respectively. Results demonstrated the qPCR was reliable for verifying porcine and bovine DNA compared to conventional PCR. Furthermore, the study concluded that the developed assay can be easily employed for the identification of porcine and bovine tissue in food products in low resource areas.

Rapid, specific and quantitative polymerase chain reaction (PCR) detection of pathogenic protozoa Entamoeba histolytica for drinking water supply

2011 ◽  
Vol 11 (4) ◽  
pp. 418-425 ◽  
Author(s):  
S. W. Lam ◽  
H. B. Zhang ◽  
L. Yu ◽  
C. H. Woo ◽  
K. N. Tiew ◽  
...  
Keyword(s):  
Real Time ◽  
Water Samples ◽  
Real Time Pcr ◽  
Genomic Dna ◽  
Tap Water ◽  
Pcr Detection ◽  
Raw Water ◽  
Conventional Pcr ◽  
Polymerase Chain ◽  
Species Specific

In this study, a quantitative species-specific polymerase chain reaction (PCR) method to rapidly detect E. histolytica in water is developed. First, the specificity of E. histolytica PCR detection was verified by using species-specific primers of 16S-like rRNA genes to clearly differentiate it from the closely related amoebae species E. dispar and E. moshkovskii. The sensitivity of this method was subsequently determined using purified E. histolytica genomic DNA and culture cells as PCR reaction templates. Results indicated that conventional PCR visualized on 1% agarose gel was able to detect as low as 0.02 pg genomic DNA and 5 cells, while real-time PCR could detect 0.01 pg genomic DNA and 2 cells of E. histolytica. The protocols for E. histolytica PCR detection in real water samples were then optimized by spiking E. histolytica cells into tap water and reservoir raw water samples. A two-round centrifugation treatment to concentrate amoeba cells directly as a PCR template was the most effective way to detect E. histolytica in spiked tap water samples, while DNA extraction after concentrating amoeba cells was required for spiked reservoir raw water samples. The detection limit of 50 E. histolytica cells in 100 ml tap water was achieved in 2 h from sample collection to real-time PCR data readout. With these established protocols, 78 tap water samples, 11 reservoir raw water samples and 4 feed water samples from Singapore water supply systems were analyzed by both conventional PCR and real-time PCR methods. No E. histolytica cell was detected in tested samples.

scholarly journals A SYBR Green real-time PCR assay to detect and quantify pork meat in processed poultry meat products

Meat Science ◽  
2013 ◽  
Vol 94 (1) ◽  
pp. 115-120 ◽  
Author(s):  
Sónia Soares ◽  
Joana S. Amaral ◽  
M. Beatriz P.P. Oliveira ◽  
Isabel Mafra
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Sybr Green ◽  
Poultry Meat ◽  
Pork Meat ◽  
Pcr Assay

scholarly journals Comparison of Real-Time PCR Quantification Methods in the Identification of Poultry Species in Meat Products

Foods ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1049
Author(s):  
Kerstin Dolch ◽  
Sabine Andrée ◽  
Fredi Schwägele
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Meleagris Gallopavo ◽  
Poultry Meat ◽  
Reference Sequence ◽  
Food Fraud ◽  
Recovery Rates ◽  
Relative Standard ◽  
Species Specific

Poultry meat is consumed worldwide and is prone to food fraud because of large price differences among meat from different poultry species. Precise and sensitive analytical methods are necessary to control poultry meat products. We chose species–specific sequences of the cytochrome b gene to develop two multiplex real-time polymerase chain reaction (real-time PCR) systems: one for chicken (Gallus gallus), guinea fowl (Numida meleagris), and pheasant (Phasianus colchicus), and one for quail (Coturnix japonica) and turkey (Meleagris gallopavo). For each species, added meat could be detected down to 0.5 % w/w. No cross reactions were seen. For these two real-time PCR systems, we applied three different quantification methods: (A) with relative standard curves, (B) with matrix-specific multiplication factors, and (C) with an internal DNA reference sequence to normalize and to control inhibition. All three quantification methods had reasonable recovery rates from 43% to 173%. Method B had more accepted recovery rates, i.e., in the range 70–130%, namely 83% compared to 75% for method A or C.

scholarly journals Detection of DNA pork in processed meat products with real-time polymerase chain reaction

Food Research ◽  
2020 ◽  
Vol 4 (5) ◽  
pp. 1719-1725
Author(s):  
W.P. Rahayu ◽  
A.R.W. Dianti ◽  
Nurjanah S. ◽  
N. Pusparini ◽  
W. Adhi
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Melting Curve ◽  
Meat Products ◽  
Processed Meat ◽  
Cyt B ◽  
Chain Reaction ◽  
Ct Value ◽  
Polymerase Chain ◽  
Cyt B Gene

Real-time polymerase chain reaction (qPCR) technique is a suitable method for identifying animal species in processed meat because of its ability to amplify a few fragments of DNA. A specific fragment of pork (mitochondrial cytochrome b gene (cytb)) was used as a DNA ladder. This study aimed to evaluate the use of a cyt-b gene generated primer for detecting the presence of pork in processed meat products by qPCR and determining the threshold cycle cut-off. The evaluation of the primer effectiveness was performed by threshold cycle (Ct) value, amplicon size by electrophoresis and melting curve. Two corned (A, B) and two jerkies (C, D) collected from the market were used as the sample. Genomic DNA from samples, fresh beef (as negative control) and fresh pork (as positive control) were extracted using Qiagen Kits. Amplification condition for 50 cycles of the cyt-b gene was performed as the initial step at 95°C for 10 mins, denaturation step at 95°C for 15 s, annealing step at 55°C for 60 s, extension step at 72°C for 30 s and post-PCR at 72°C for 3 mins. The threshold cycle (Ct) cut-off less than 30 confirmed as pork positive. The result obtained indicated that sample A and D were pork negative, with Ct value respectively 40.73 and 43.59. Melting temperatures of amplicon were ranged from 79.5 to 80.5°C, only differed by 1°C, and the amplicon electrophoresis resulting in a single band of the same size (149 bp). Hence, the melting curve analysis and electrophoresis of PCR products were not able to differentiate between pork and beef.

EVALUATION OF THE QUALITY OF MEASURING PROCESSES IN THE MANUFACTURE OF POULTRY MEAT PRODUCTS

2019 ◽  
pp. 33-40
Author(s):  
O. A. LEONOV ◽  
◽  
N. J. SHKARUBA ◽  
A. A. ODINTSOVA ◽  
◽  
...  
Keyword(s):  
Meat Products ◽  
Poultry Meat

Ukraine’s foreign trade in meat and meat products: trends and geographical aspects

2020 ◽  
pp. 48-52
Author(s):  
Olena Lyakhovska
Keyword(s):  
Small Businesses ◽  
Structural Changes ◽  
Raw Materials ◽  
Meat Products ◽  
World Market ◽  
Poultry Meat ◽  
Positive Development ◽  
Meat Production ◽  
Meat Industry ◽  
Geographical Structure

Current trends in meat and meat products exports are considered. The dynamics and structural changes in meat exports by types and degree of processing (beef and veal, pork, poultry, finished meat products) are analyzed. The paper proves that in recent years, only meat and edible poultry by-products have taken a competitive position in the world market. Accordingly, the dynamics of exports of this type of meat is positive, and the share of these products in the structure of exported meat is the largest (86.09%). Possible threats to the growth of import dependence on certain types of meat products, in particular, chilled and frozen beef and veal, pork, are indicated. The rating of the largest countries-importers of meat and meat products (by types) from Ukraine is built, their share in the structure of meat exports is determined. Changes in the geographical structure of exports have been studied, which indicate a low level of diversification of the geography of exports of certain types of meat (pork, beef). At the same time, a positive development of the geographical structure of poultry meat exports (exported to more than 100 countries) was revealed. Problems of the export potential development in the meat industry of Ukraine are revealed. In particular, the problems of livestock development (outdated technologies and production techniques that reduce the efficiency and productivity; inefficiency of the organization of purchase and processing of meat, which leads to reduced breeding in small businesses and households; inefficiency of state incentives, etc.) and industrial meat production (lack of raw materials, obsolescence of fixed assets and production technologies, poorly developed logistics, etc.). The solution of these problems requires: a systematic approach, which will encourage the cooperation of agricultural and industrial production, cooperation with importers of meat products and protection of domestic producers in the domestic and international markets, creating a favorable investment and information environment that will promote the meat industry development.

Оrganic Meat Production of Broiler Chickens Hubbard Redbro Cross

2020 ◽  
Keyword(s):  
Broiler Chickens ◽  
Raw Materials ◽  
Animal Health ◽  
Meat Products ◽  
Poultry Meat ◽  
Meat Production ◽  
Significant Difference ◽  
Environmentally Safe ◽  
The Impact

This article presents the results of studying the impact of housing and feeding conditions on broiler chickens of Hubbard RedBro cross, as well as the quality of products obtained when using floor and cage content, in a farm. It established that when receiving a mixed feed of own production using feed raw materials grown on a farm without the use of pesticides, a statistically significant decrease in potentially dangerous substances for animal health is recorded. Compared with factory feed, it has reduced the content of pesticides by 14 times, and mercury and arsenic by 24 times, cadmium by five times, and lead by ten times. The results of the study of economic indicators of growing Hubbard RedBro cross broiler chickens, as well as the chemical composition and quality of carcasses, indicated that there was no significant difference between the floor and cell conditions of keeping. Still, the use of a diet based on eco-feeds contributed to a statistically significant decrease in the concentration of toxic metals in the muscles of the poultry of the experimental groups. As a result, it found that the use of the studied compound feed in the diets of broiler chickens increased the indicators of Biosafety and ensured the production of environmentally safe ("organic") poultry meat products.

scholarly journals Effect of Herbal Addition on the Microbiological, Oxidative Stability and Sensory Quality of Minced Poultry Meat

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1537
Author(s):  
Danuta Jaworska ◽  
Elżbieta Rosiak ◽  
Eliza Kostyra ◽  
Katarzyna Jaszczyk ◽  
Monika Wroniszewska ◽  
...  
Keyword(s):  
Oxidative Stability ◽  
Sensory Quality ◽  
Meat Products ◽  
Black Pepper ◽  
Poultry Meat ◽  
Piper Nigrum ◽  
Thymus Vulgaris ◽  
Processing And Storage ◽  
And Storage

The study aimed to assess the effect of herbal additions with antioxidant properties (pepper, thyme and oregano) on the microbiological and oxidative stability as well as the sensory quality of minced poultry meat. Meatballs treatments without additives and treatments with the addition of three types of spices in two forms—dry spices and industrial extracts were examined. Popular seasoning additives of oregano (Origanum vulgare), thyme (Thymus vulgaris) and black pepper (Piper nigrum) at 0.3% of dry herbal or 0.003% as industrial extract were added to minced meat. The microbiological, chemical and sensory tests were performed at specified times and storage temperature. Based on the studied criteria, products maintained constant and adequate quality by up to 10 days while stored in 4 °C. In the case of all tested samples, the overall sensory quality began to deteriorate after 10 days of storage. The preservative role of herbs and extracts in meat products during processing and storage was observed. Oregano and black pepper in both forms maintained good microbial quality and showed their inhibitory effects on the growth of psychrotrophic bacteria. It was observed that dried herbs revealed a stronger antioxidant effect than additives in the form of extracts. The studied dried herbs played an antioxidant, antimicrobial and preservative role in meat products during processing and storage.

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