Specific PCR Detection of Tiger, Leopard, and Lion Ingredients from Test Samples

Abstract A PCR method was developed for specifc detection of tiger, leopard, and lion DNA from test specimens for inspection and quarantine or for law-enforced animal protection. Three pairs of specifc primers were designed based on the mitochondrial cytochrome b gene of tiger, leopard, and lion and used in the PCR testing. To mimic the effect of food processing on the sensitivity of the test, the tiger muscle and bovine bonemeal powder samples were treated at 133°C for 30 min. At this processing condition, the method was sensitive enough to detect as low as 0.05% of tiger-derived ingredients from the mixed bonemeal powders. The data demonstrate that our PCR method is convenient and economic, with high sensitivity and repeatability, and can be used to detect and identify tiger, leopard, and lion ingredients from various test samples.

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Species Identification and Quantification of Bovine Bone Marrow from Non-Bovine Products using DNA and Protein Fingerprints

10.21203/rs.3.rs-1062457/v1 ◽

2021 ◽

Author(s):

Yize Guan ◽

Nan Li ◽

Tancheng Li ◽

Liyuan Sun ◽

Li Ming Cheng

Keyword(s):

Bone Marrow ◽

Multiplex Pcr ◽

High Sensitivity ◽

Bovine Bone ◽

Assay Sensitivity ◽

Conserved Sequence ◽

Specific Pcr ◽

Modified Method ◽

Sds Page ◽

Mitochondrial Cytochrome B

Abstract Bovine bone marrow is traditionally regarded as a highly nutritious food that has been widely used as a medicinal and healthy food for several decades in China. A large number of adulterated and counterfeit bone marrows from pigs and donkeys have been used in place of bovine bone marrow in commercial products, which are almost identical morphologically to species. Therefore, we explored a feasibility of fingerprinting of multiplex PCR gels combined with imaged SDS-PAGE gels of proteins. Three pairs of specific primers for bovine, pig and donkey were designed according to the conserved sequence in mitochondrial cytochrome b. The optimal reaction conditions for triple PCR were optimized. A modified method was used to extract total proteins and SDS-PAGE gels of proteins were set up. A three-fold PCR detection assay was successfully established to identify three species of bovine, pig and donkey. Three primers have good specificity and high sensitivity. Additionally, the assay sensitivity test confirmed that the extracted DNA concentration was the lowest of bovine bone marrow at 0.1 pg. The assay also showed 100% specificity. The electrophoretogram of SDS-PAGE showed distinct electrophoresis profiles among three species. The established fingerprint of multiplex PCR DNA gels combined with imaged SDS-PAGE gels of proteins has strong specificity and can be quickly and accurately used for the identification of bovine bone marrow. Rapid authentication of bovine bone marrow and differentiation from non-bovine products can be achieved using an improved SDS alkali denaturation method, species-specific PCR assay combined with imaged SDS-PAGE gels of proteins.

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Detection of Legionella Species in Respiratory Specimens Using PCR with Sequencing Confirmation

Journal of Clinical Microbiology ◽

10.1128/jcm.38.5.1709-1712.2000 ◽

2000 ◽

Vol 38 (5) ◽

pp. 1709-1712 ◽

Cited By ~ 92

Author(s):

J. L. Cloud ◽

K. C. Carroll ◽

P. Pixton ◽

M. Erali ◽

D. R. Hillyard

Keyword(s):

Legionella Pneumophila ◽

High Sensitivity ◽

High Specificity ◽

Rrna Gene ◽

Specific Pcr ◽

Pcr Method ◽

Specific Pcr Assay ◽

Clinically Significant ◽

Tract Infections ◽

Culture Negative

Legionella spp. are a common cause of community-acquired respiratory tract infections and an occasional cause of nosocomial pneumonia. A PCR method for the detection of legionellae in respiratory samples was evaluated and was compared to culture. The procedure can be performed in 6 to 8 h with a commercially available DNA extraction kit (Qiagen, Valencia, Calif.) and by PCR with gel detection. PCR is performed with primers previously determined to amplify a 386-bp product within the 16S rRNA gene of Legionella pneumophila. We can specifically detect the clinically significant Legionella species including L. pneumophila, L. micdadei, L. longbeachae, L. bozemanii, L. feeleii, and L. dumoffii. The assay detects 10 fg (approximately two organisms) of legionella DNA in each PCR. Of 212 clinical specimens examined by culture, 100% of the culture-positive samples (31 of 31) were positive by this assay. By gel detection of amplification products, 12 of 181 culture-negative samples were positive forLegionella species by PCR, resulting in 93% specificity. Four of the 12 samples with discrepant results (culture negative, PCR positive) were confirmed to be positive for Legionellaspecies by sequencing of the amplicons. The legionella-specific PCR assay that is described demonstrates high sensitivity and high specificity for routine detection of legionellae in respiratory samples.

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PCR Detection of Alternaria spp. in Processed Foods, Based on the Internal Transcribed Spacer Genetic Marker

Journal of Food Protection ◽

10.4315/0362-028x.jfp-10-110 ◽

2011 ◽

Vol 74 (2) ◽

pp. 240-247 ◽

Cited By ~ 16

Author(s):

MIGUELÁNGEL PAVÓN ◽

ISABEL GONZÁLEZ ◽

MARÍA ROJAS ◽

NICOLETTE PEGELS ◽

ROSARIO MARTÍN ◽

...

Keyword(s):

Food Processing ◽

Internal Transcribed Spacer ◽

Rapid Identification ◽

Food Samples ◽

Pcr Analysis ◽

Infant Food ◽

Rrna Gene ◽

Tomato Pulp ◽

Pcr Method ◽

Fungal Contaminants

The genus Alternaria is considered one of the most important fungal contaminants of vegetables, fruits, and cereals, producing several mycotoxins that can withstand food processing methods. Conventional methods for Alternaria identification and enumeration are laborious and time-consuming, and they might not detect toxigenic molds inactivated by food processing. In this study, a PCR method has been developed for the rapid identification of Alternaria spp. DNA in foodstuffs, based on oligonucleotide primers targeting the internal transcribed spacer (ITS) 1 and ITS2 regions of the rRNA gene. The specificity of the Alternaria-specific primer pair designed (Dir1ITSAlt–Inv1ITSAlt) was verified by PCR analysis of DNA from various Alternaria spp., and also from several fungal, bacterial, yeast, animal, and plant species. The detection limit of the method was 102 CFU/ml in viable culture, heated culture, or experimentally inoculated tomato pulp. The applicability of the method for detection of Alternaria spp. DNA in foodstuffs was assessed by testing several commercial samples. Alternaria DNA was detected in 100% of spoiled tomato samples, 8% of tomato products, and 36.4% of cereal-based infant food samples analyzed.

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Effects of thermal processing on the allergenicity, structure, and critical epitope amino acids of crab tropomyosin

Food & Function ◽

10.1039/d0fo02869j ◽

2021 ◽

Author(s):

Meng Liu ◽

Tian-Jiao Han ◽

Fei Huan ◽

Meng-Si Li ◽

Fei Xia ◽

...

Keyword(s):

Amino Acids ◽

Food Processing ◽

Thermal Processing ◽

Effect Of Food

Food processing can change the structure and immunoreactivity of purified allergens, but the effect of food processing on the immunoreactivity of the processed and purified allergen is still poorly understood.

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Effect of food processing on the antioxidant activity of flavones from Polygonatum odoratum (Mill.) Druce

Open Life Sciences ◽

10.1515/biol-2021-0010 ◽

2021 ◽

Vol 16 (1) ◽

pp. 92-101

Author(s):

Guanghui Xia ◽

Xinhua Li ◽

Zhen Zhang ◽

Yuhang Jiang

Keyword(s):

Antioxidant Activity ◽

High Pressure ◽

Food Processing ◽

Yeast Fermentation ◽

Pressure Treatment ◽

Processing Methods ◽

High Pressure Treatment ◽

Effect Of Food ◽

The Stability

Abstract Polygonatum odoratum (Mill.) Druce (POD) is a natural plant widely used for food and medicine, thanks to its rich content of a strong antioxidant agent called homoisoflavones. However, food processing methods could affect the stability of POD flavones, resulting in changes to their antioxidant activity. This study attempts to evaluate the antioxidant activity of POD flavones subject to different processing methods and determines which method could preserve the antioxidant activity of POD flavones. Therefore, flavones were extracted from POD samples, which had been treated separately with one of the four processing methods: extrusion, baking, high-pressure treatment, and yeast fermentation. After that, the antioxidant activity of the flavones was subject to in vivo tests in zebrafish embryos. The results show that yeast fermentation had the least disruption to the antioxidant activity of POD flavones, making it the most suitable food processing method for POD. By contrast, extrusion and high-pressure treatment both slightly weakened the antioxidant activity of the flavones and should be avoided in food processing. The research results provide a reference for the development and utilization of POD and the protection of its biological activity.

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

Applied and Environmental Microbiology ◽

10.1128/aem.69.12.7430-7434.2003 ◽

2003 ◽

Vol 69 (12) ◽

pp. 7430-7434 ◽

Cited By ~ 77

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.

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The effect of food processing on the amount of trace elements and their bioavailability: a review

Journal of Food Safety and Hygiene ◽

10.18502/jfsh.v6i2.6520 ◽

2021 ◽

Author(s):

Maral Neyestani ◽

Parisa Shavali Gilani ◽

Mohadeseh Fesahat ◽

Ebrahim Molaee-Aghaee ◽

Nabi Shariatifar

Keyword(s):

Trace Elements ◽

Trace Metals ◽

Food Processing ◽

Food Production ◽

Food Additives ◽

Food Product ◽

Special Method ◽

Effect Of Food ◽

Almost All ◽

Main Factor

Trace elements are compounds that are essential in small amounts for biochemical reactions and to maintain human health. Almost all foods can contain varying amounts of these metals. In this study, the effects of food processing on the content of trace metals are investigated. Extensive interpretations of processing, including aspects of food production and specific examples of changes in metal content due to processing will be discussed. Pre-consumption food processing to improve rheological properties and increase shelf life is inevitable, which changes the bioavailability and amount of these compounds in different directions depending on the process. The amount of these trace metals in the food product can be affected by various conditions such as heating, fermentation, food additives, etc. The main factor in reducing trace elements in food, especially the use of heat in a special method and on the other hand, factors such as fermentation can also increase the bioavailability of these elements.

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Prevalence and Serotype Determination of Streptococcus agalactiae Isolated From Non-Pregnant Women in Tehran, Iran

ACTA MEDICA IRANICA ◽

10.18502/acta.v57i9.2641 ◽

2020 ◽

Author(s):

Leila Goudarzi ◽

Mohammad Bagher Khalili ◽

Mahmood Vakili ◽

Maryam Sadeh

Keyword(s):

Pregnant Women ◽

Streptococcus Agalactiae ◽

Group B Streptococcus ◽

Cross Sectional Study ◽

Chi Square ◽

Cross Sectional ◽

Specific Pcr ◽

Pcr Method ◽

Group B ◽

Tehran Area

Consequence of Streptococcus agalactiae, Group B Streptococcus (GBS) relating infant’s diseases are well documented. Although many women carry this bacterium in their vagina, they may transfer to their infant during delivery and may result in different neonatal invasive diseases. The aim of this study was to determine the prevalence of GBS and serotyping the isolated species among un-selective non-pregnant women who attended two gynecology clinics in Tehran. In this cross-sectional study, a total of 560 vaginal samples collected from non-pregnant women. Following inoculation of the specimen on Blood Agar, the standard technology was applied for the final identification of GBS. Detected GBS species were further confirmed using specific PCR directed on dlts gene. Capsular serotyping was done by using the multiplex PCR method. The chi-square method was used for statistical analysis. Fifty (8.9%) out of 560 non-pregnant women were carriers of GBS. The most common types were III (36%), followed by type II (32%), Ia (26%), and Ib (6%), respectively. Results represent that the prevalence rate of GBS in non-pregnant women was reliable and similar to what obtained from pregnant women. In addition, the serotype III was found the most dominant types, as well as other investigations in the Tehran area. Therefore, vaccine designation based on type III is recommended.

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