scholarly journals Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat

2001 ◽  
Vol 43 (5) ◽  
pp. 247-250 ◽  
Author(s):  
Luciana Ruschel dos SANTOS ◽  
Vladimir Pinheiro do NASCIMENTO ◽  
Sílvia Dias de OLIVEIRA ◽  
Maristela Lovato FLORES ◽  
Alexandre Pontes PONTES ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Thermal Treatment ◽  
Dna Extraction ◽  
Salmonella Enteritidis ◽  
Chicken Meat ◽  
Chain Reaction ◽  
Initial Dilution ◽  
Time Period ◽  
Polymerase Chain ◽  
Meat Samples

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10-7, 10-8 or 10-9 CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10-9 CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.

scholarly journals Morphological, Biochemical and Genotypic Analysis of Zoonotic Campylobacter jejuni Isolated from Chicken Meat Samples

2022 ◽  
Author(s):  
Sumedha Bobade ◽  
K. Vijayarani ◽  
K.G. Tirumurugaan ◽  
A. Thangavelu ◽  
S. Vairamuthu
Keyword(s):  
Polymerase Chain Reaction ◽  
Campylobacter Jejuni ◽  
Chicken Meat ◽  
Poultry Meat ◽  
Phenotypic Characterization ◽  
Meat Production ◽  
Chain Reaction ◽  
Genotypic Analysis ◽  
Polymerase Chain ◽  
Meat Samples

Background: Campylobacter species are a leading cause of most important food-borne diarrhoeal illness worldwide while, poultry has been identified as a significant cause of Campylobacter infection in humans. C. jejuni is highly effective in colonizing chicken intestinal mucosa without causing any clinical manifestations and the consumption of poultry meat is the major source of transmission of bacteria to humans. Methods: The total of 19 chicken meat samples collected from retail markets in Chennai were screened by cultural examination, further subjected to phenotypic characterization using biochemical test and genotypic characterization using polymerase chain reaction assay targeting hip O and map A genes. Result: All the isolates showed growth on modified blood free charcoal cefoperazone deoxycholate agar media (mCCDA) and 18 (94.73%) samples showed typical morphological characteristics. The 12 (63.15%) isolates showed biochemical reactions positive. The results from polymerase chain reaction showed that 10 (83.33%) isolates were positive for C. jejuni. This study suggested that, it is essential to investigate the incidence of Campylobacter jejuni infection in poultry and the risk factors at all production stages of meat production to help reducing the disease in humans in terms of food safety.

scholarly journals Prevalence and diversity of Arcobacter spp. in retail chicken meat in Turkey

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Celenk Molva ◽  
Halil Ibrahim Atabay
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Chicken Meat ◽  
Species Level ◽  
Waterborne Pathogens ◽  
Chain Reaction ◽  
Retail Markets ◽  
Specific Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Retail Chicken Meat

Arcobacters are food and waterborne pathogens associated with human and animal infections. The objective of the present study was to investigate the prevalence and diversity of <em>Arcobacter</em> spp. in commercially sold chicken meat in İzmir region of Turkey. For this purpose, 100 samples including legs (n=40), 17 chicken quarters (n=17), drumstickers (n=16), breasts (n=11), wings (n=10), and carcasses (n=6) were collected from different retail markets. A total of 65 isolates were confirmed as <em>Arcobacter</em> spp. from 55 samples by genus-specific polymerase chain reaction (PCR). The prevalence of <em>Arcobacter</em> spp. was 32.5, 81.3, 64.7, 72.7, 83.3, and 50% for legs, drumstickers, chicken quarters, breasts, carcasses and wings, respectively. Based on the multiplex-PCR, most of the isolates were identified as <em>A. butzleri</em> (n=45, 80%), followed by <em>A. cryaerophilus</em> (n=2, 3.6%), <em>A. skirrowii</em> (n=1, 1.8%) and 17 isolates (30.9%) could not be identified at the species level.

scholarly journals Method Comparison of DNA Isolation and Quantification for Fish and Seafood Authenticity Determination

2018 ◽  
Vol 13 (3) ◽  
pp. 115
Author(s):  
Dwiyitno Dwiyitno ◽  
Stefan Hoffman ◽  
Koen Parmentier ◽  
Chris Van Keer
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Dna Isolation ◽  
Blue Mussel ◽  
Pcr Amplification ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Seafood Products ◽  
Polymerase Chain ◽  
Commercial Kit

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen

scholarly journals Longest reported case of symptomatic COVID-19 reporting positive for over 230 days in an immunocompromised patient in the United States

2021 ◽  
Vol 9 ◽  
pp. 2050313X2110400
Author(s):  
Bilal Chaudhry ◽  
Lidiya Didenko ◽  
Maaria Chaudhry ◽  
Andrew Malek ◽  
Kirill Alekseyev
Keyword(s):  
Risk Factors ◽  
Polymerase Chain Reaction ◽  
Immunocompromised Patient ◽  
The United States ◽  
Immunocompromised Patients ◽  
Chain Reaction ◽  
Viral Shedding ◽  
Time Period ◽  
Increased Risk ◽  
Polymerase Chain

Coronavirus 2019 (COVID-19) pneumonia was first noted in Wuhan, China. Since the start of the pandemic, there have been millions of cases diagnosed. The average time from onset of symptoms to testing negative SARS-CoV-2 via reverse transcription polymerase chain reaction is roughly 25 days. In patients who continually test positive for COVID-19, it is essential to determine precisely which risk factors contribute to the increase in viral shedding duration. We present a case about a 62-year-old man who has persistently tested positive for COVID-19 for more than 230 days. We followed his treatment course, in which he had been hospitalized multiple times since the onset of symptoms back in April 2020. We have determined that patients with immunosuppression, especially those taking corticosteroids, are at increased risk of prolonged viral shedding. It is essential to continually monitor these immunocompromised patients as they required a greater time period in order to have an appropriate immune response in which antibodies are created.

scholarly journals Molecular diagnosis of the curly lettuce parasitic contamination from hydroponic cultivation from supermarkets

2020 ◽  
Vol 29 (4) ◽  
Author(s):  
Fernanda Pinto-Ferreira ◽  
Jonatan Batista Reis ◽  
Aline Ticiani Pereira Paschoal ◽  
Letícia Santos Balbino ◽  
Amanda Bertão-Santos ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Dna Extraction ◽  
Chain Reaction ◽  
Cryptosporidium Spp ◽  
Hydroponic Cultivation ◽  
Polymerase Chain ◽  
Fluctuation Method ◽  
The City ◽  
Commercial Kit

Abstract The consumption of vegetables has increased in recent years due to the search for a healthier diet that is rich in fiber and has fewer calories. To assess the parasitic contamination of lettuce sold in markets, a survey of parasites was carried out from a supermarket chain in the city of Londrina, Paraná. A total of thirty samples of lettuce were purchased in the ten markets visited, three in each, of which ten were conventionally cultivated, ten were hydroponically cultivated, and ten were organically cultivated. All samples were analyzed using the sedimentation methods of Hoffman, Pons and Janer and the fluctuation method of Faust and colleagues and Willis with adaptations. In addition, the samples were subjected to DNA extraction by a commercial kit and polymerase chain reaction to detect Toxoplasma gondii, Cryptosporidium spp. and Giardia spp., which are protozoa that cause food and waterborne parasitic outbreaks. All samples were negative for sedimentation and flotation techniques. One of the hydroponically cultivated samples was positive for T. gondii. The results demonstrate the risk of curly lettuce contamination from hydroponic cultivation and the need for proper cleaning of these foods before consumption.

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

Sex identification of pigeons using polymerase chain reaction analysis with simple DNA extraction

2019 ◽  
Vol 12 (2) ◽  
pp. 45-48
Author(s):  
Shao-jie Liang ◽  
Ming-xia Chen ◽  
Chun-qi Gao ◽  
Hui-chao Yan ◽  
Guo-long Zhang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Polymerase Chain Reaction Analysis ◽  
Sex Identification ◽  
Z Chromosome ◽  
W Chromosome ◽  
Chain Reaction ◽  
Assay Time ◽  
Polymerase Chain ◽  
The Cost

Sex identification plays an important role in avian production. Hitherto, it is difficult to distinguish the sexes of monomorphic birds based on their external features. The chromo-helicase-DNA-binding genes contain CHD-W gene and CHD-Z gene, which are located on the W chromosome and Z chromosome, respectively. Since CHD-W gene is unique to females, the polymerase chain reaction can be used for sex identification. However, extracting DNA procedures for verifying the sex is tedious and expensive. To address these disadvantages, the objective of this study was to develop a simple DNA extraction assay to efficiently process blood, liver, and feather samples. The results showed that 2% dimethylsulfoxide was suitable for processing blood, and phosphate-buffered saline was suitable for processing liver and feather samples. The specific primers were designed, and the length of the targets is 474 bp on Z chromosome and 319 bp on W chromosome. The pigeons were identified as females based on the presence of two bands on the gel, and as males based on the presence of one band. Taken together, our results suggested that feather samples were more appropriate than blood or liver for sex identification of pigeons. Compared to the traditional DNA extraction, this method shortened the assay time and reduced the cost.

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