scholarly journals Methods of mycobacterial DNA isolation from different biological material: a review

2012 ◽  
Vol 51 (No. 5) ◽  
pp. 180-192 ◽  
Author(s):  
J. Hosek ◽  
P. Svastova ◽  
M. Moravkova ◽  
I. Pavlik ◽  
M. Bartos
Keyword(s):  
Dna Isolation ◽  
Economic Losses ◽  
Human Beings ◽  
Chain Reaction ◽  
Serious Infections ◽  
Biochemical Characterisation ◽  
Long Time ◽  
Polymerase Chain ◽  
Wide Family

Mycobacteria cause serious infections in animals and human beings. Huge economic losses on farms are caused by selected species of this wide family. A high risk of transmission of infection from animal to human exists. The knowledge of exact pathogen characteristics is an important factor which can improve quick and adequate healing. Cultivation and determination of phenotype is still the “gold standard”, but has the disadvantage of taking a long time and also low detection limit. Biochemical characterisation of isolates is not exact, and it is expensive. A more popular method used is the amplification of specific loci by polymerase chain reaction (PCR). For this method, the isolation of sufficient amounts of purified DNA is necessary. In this paper the most frequently used method for DNA isolation from live mycobacterial cells, body fluids, tissues, histological samples and forensic materials are outlined. This paper assists only as guide for these methods, so we describe them briefly.

scholarly journals Overview of Safety Measures at Selected Airports during the COVID-19 Pandemic

2021 ◽  
Vol 13 (15) ◽  
pp. 8499
Author(s):  
Monika Blišťanová ◽  
Michaela Tirpáková ◽  
Ľubomíra Brůnová
Keyword(s):  
Time Frame ◽  
Organizational Changes ◽  
Rt Pcr ◽  
Final Phase ◽  
Chain Reaction ◽  
Scoring Method ◽  
Safety Measures ◽  
Long Time ◽  
Polymerase Chain ◽  
Cleaning And Disinfection

The year 2020 was very challenging for the whole world, given the outbreak of the ongoing coronavirus-related pandemic, and was marked in particular by overcoming new hitherto unknown obstacles. For air transport, in particular, airlines stopped flying altogether and were forced to ground hundreds of planes worldwide involuntarily. Airports had to close their terminals for a long time, wholly suspend operations, and its resumption required significant organizational changes. This article summarizes the measures related to the COVID-19 pandemic adopted by airports to minimize the risk of spreading the disease. The article focuses on countermeasures and their implementation at selected airports in a specific time frame and airports’ behavior during a pandemic which varies depending on country and time of the year. The results demonstrated that steps being taken at airports include the use of face coverings or masks, social distance, enhanced cleaning and disinfection, or temperature checks and/or symptoms (fever, loss of smell, chills, cough, shortness of breath), RT-PCR (reverse transcription-polymerase chain reaction) screening and data collection with health declaration. These measures have now become an essential standard for the operation of airports and can, therefore, be used to assess the level of airport safety achieved. In the final phase, the article evaluates the level of achieved airport safety based on the proposed scoring method.

scholarly journals Acceptance of a Symptom-Based Approach for COVID-19 rtRT-PCR Testing to Conserve Resources in a Lower Middle-Income Setting

2021 ◽  
Vol 12 ◽  
pp. 215013272098771
Author(s):  
S. M. Rashed Ul Islam ◽  
Tahmina Akther ◽  
Md. Abdullah Omar Nasif ◽  
Sharmin Sultana ◽  
Saif Ullah Munshi
Keyword(s):  
Primary Method ◽  
Chain Reaction ◽  
Middle Income ◽  
Clinical Recovery ◽  
Clinical Presentations ◽  
Long Time ◽  
Nasopharyngeal Secretion ◽  
Polymerase Chain ◽  
Set Up

SARS-CoV-2 initially emerged in Wuhan, China in late 2019. It has since been recognized as a pandemic and has led to great social and economic disruption globally. The Reverse Transcriptase Real-Time Polymerase Chain Reaction (rtRT-PCR) has become the primary method for COVID-19 testing worldwide. The method requires a specialized laboratory set up. Long-term persistence of SARS-CoV-2 RNA in nasopharyngeal secretion after full clinical recovery of the patient is regularly observed nowadays. This forces the patients to spend a longer period in isolation and test repeatedly to obtain evidence of viral clearance. Repeated COVID-19 testing in asymptomatic or mildly symptomatic cases often leads to extra workload for laboratories that are already struggling with a high specimen turnover. Here, we present 5 purposively selected cases with different patterns of clinical presentations in which nasopharyngeal shedding of SARS-CoV-2 RNA was observed in patients for a long time. From these case studies, we emphasized the adoption of a symptom-based approach for discontinuing transmission-based precautions over a test-based strategy to reduce the time spent by asymptomatic and mildly symptomatic COVID-19 patients in isolation. A symptom-based approach will also help reduce laboratory burden for COVID-19 testing as well as conserve valuable resources and supplies utilized for rtRT-PCR testing in an emerging lower-middle-income setting. Most importantly, it will also make room for critically ill COVID-19 patients to visit or avail COVID-19 testing at their convenience.

Evaluation of polymerase chain reaction and DNA isolation protocols for detection of genetically modified soybean

2007 ◽  
Vol 42 (10) ◽  
pp. 1249-1255 ◽  
Author(s):  
Cibele dos Santos Ferrari ◽  
Luciana Lehmkuhl Valente ◽  
Fábio Cristiano Angonesi Brod ◽  
Caroline Tagliari ◽  
Ernani Sebastião Sant'Anna ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Isolation ◽  
Genetically Modified ◽  
Chain Reaction ◽  
Genetically Modified Soybean ◽  
Polymerase Chain

DNA Isolation from Recalcitrant Materials Such as Tree Roots, Bark, and Forest Soil for the Detection of Fungal Pathogens by Polymerase Chain Reaction

1998 ◽  
Vol 262 (1) ◽  
pp. 79-82 ◽  
Author(s):  
Günther Bahnweg ◽  
Steffen Schulze ◽  
Evelyn M. Möller ◽  
Hilkea Rosenbrock ◽  
Christian Langebartels ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Forest Soil ◽  
Fungal Pathogens ◽  
Dna Isolation ◽  
Chain Reaction ◽  
Tree Roots ◽  
Polymerase Chain

Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocks

2012 ◽  
Vol 15 (2) ◽  
pp. 337-344 ◽  
Author(s):  
D. Roussan ◽  
I. Shaheen ◽  
G. Khawaldeh ◽  
W. Totanji ◽  
R. Al-Rifai
Keyword(s):  
Polymerase Chain Reaction ◽  
Broiler Chicken ◽  
Simultaneous Detection ◽  
Adenovirus Type ◽  
Economic Losses ◽  
Type I ◽  
Chain Reaction ◽  
Group I ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Simultaneous detection of astrovirus, rotavirus, reovirus and adenovirus type I in broiler chicken flocksEnteric diseases cause substantial economic losses to the poultry industry. Astroviruses, rotaviruses, reoviruses, and adenovirus type 1 have been reported as a significant cause of intestinal symptoms in poultry. In the present study, intestinal samples from 70 commercial broiler chicken flocks were examined for the presence of astroviruses, rotavirus, and reovirus by reverse transcription-polymerase chain reaction, and for the presence of group I adenovirus by polymerase chain reaction. Astroviruses were identified in 38.6% of samples tested. Both avian nephritis virus and chicken astrovirus were identified in the astrovirus positive flocks, where 74.1% of these flocks were positive for only one type of astrovirus, whereas, 25.9% of these flocks were positive for both types of astrovirus. Reoviruses, rotaviruses, and adenoviruses were identified in 21.4, 18.6, and 14.3% of these flocks, respectively. Concomitant infection with two or more viruses in the same flock were also prominent, where 5.7, 5.7, 2.9, 2.9, 1.4, and 1.4% of these flocks were positive with both astrovirus and rotavirus; astrovirus and adenovirus; astrovirus and reovirus; rotavirus and adenovirus; rotavirus and reovirus; and reovirus and adenovirus respectively. Moreover, 4.3 and 2.7% of these flocks were positive for astrovirus, reovirus, and adenovirus; and astrovirus, reovirus, and rotavirus, respectively. Further studies will focus on identifying specific viral factors or subtypes/subgroups associated with disease through pathogenesis studies, economic losses caused by infections and co-infections of these pathogens, and the costs and benefits of countermeasures.

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

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