scholarly journals Fast Identification of Yersinia pestis, Bacillus anthracis and Francisella tularensis Based on Conventional PCR

2013 ◽  
Vol 62 (4) ◽  
pp. 453-455 ◽  
Author(s):  
ALEKSANDRA A. ZASADA ◽  
KAMILA FORMIŃSKA ◽  
KATARZYNA ZACHARCZUK
Keyword(s):  
Public Health ◽  
Real Time ◽  
Francisella Tularensis ◽  
Real Time Pcr ◽  
Health Impact ◽  
Diagnostic Tools ◽  
Conventional Pcr ◽  
The Public ◽  
Detection And Identification ◽  
Appropriate Treatment

Rapid and accurate diagnostic tools for detection and identification of Y pestis, B. anthracis and F. tularensis are essential for timely initial appropriate treatment of exposed individuals, which will be critical to their survival, as well as for reduction of the public health impact and the spread of the disease. The paper presents application of fast polymerases and fast dry electrophoresis in conventional PCR as an alternative for real-time PCR application for detection and identification of the above pathogens. The proposed method takes less than 50 min. to obtain final results of the tests and is cheaper than real-time PCR.

scholarly journals Comparing the efficacy of three diagnostic tools (microscopic examination, conventional PCR, and Real-Time PCR) in detecting Theileria equi infection among Egyptian equines

2021 ◽  
Author(s):  
Ahmed M. Soliman ◽  
Moaz M. Amer ◽  
Fareed Uddin Memon
Keyword(s):  
Real Time ◽  
Microscopic Examination ◽  
Real Time Pcr ◽  
Protozoan Parasite ◽  
Diagnostic Tools ◽  
Microscopical Examination ◽  
Conventional Pcr ◽  
Theileria Equi ◽  
Diagnostic Assays ◽  
Blood Smears

Abstract Equine theileriosis represents one of the main and serious health problems affecting equines industry globally, that is caused by tick-borne protozoan parasite called T. equi. This study aimed to assess the sensitivity of three diagnostic tools named: microscopic examination of a blood smear, conventional PCR, and Real-Time PCR (qPCR) to detect T. equi among equine population (n = 116) raised in Giza Governorate, Egypt. Microscopic examination of Giemsa-stained blood smears revealed the infection of 16.4% (19/116) of examined equines by T. equi while conventional PCR and qPCR revealed that 29.3% (34/116) and 43.1% (50/116) of examined equines were infected with T. equi respectively. Our results demonstrated that the qPCR had the highest sensitivity (100%) followed by conventional PCR (68%) while microscopic examination had the lowest sensitivity (38%). Furthermore, the negative predictive value (NPV) of qPCR was the highest (100%) compared to conventional PCR and microscopical examination (80.49% and 68.04% respectively) which revealed that all negative cases detected by qPCR were certainly correct compared to the other two diagnostic assays. Therefore, it is highly recommended to incorporate PCR diagnostic assays (conventional PCR and qPCR) alongside microscopic examination to evaluate the epidemiological status of equine theileriosis.

scholarly journals Nanopore sequencing for the detection and the identification of Xylella fastidiosa subspecies and sequence types from naturally infected plant material

2019 ◽  
Author(s):  
Luigi Faino ◽  
Valeria Scala ◽  
Alessio Albanese ◽  
Vanessa Modesti ◽  
Alessandro Grottoli ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Plant Material ◽  
Xylella Fastidiosa ◽  
Infected Plant ◽  
Diagnostic Tools ◽  
Nanopore Sequencing ◽  
Direct Dna Sequencing ◽  
Detection And Identification ◽  
Infected Plant Material

SummaryXylella fastidiosa (Xf) is a polyphagous gram-negative bacterial plant pathogen that can infect more than 300 plant species. It is endemic in America while, in 2013, Xf subsp. pauca was for the first time reported in Europe on olive tree in the Southern Italy. The availability of fast and reliable diagnostic tools is indispensable for managing current and future outbreaks of Xf.In this work, we used the Oxford Nanopore Technologies (ONT) device MinION platform for detecting and identifying Xf at species, subspecies and Sequence Type (ST) level straight from infected plant material. The study showed the possibility to detect Xf by direct DNA sequencing and identify the subspecies in highly infected samples. In order to improve sensitivity, Nanopore amplicon sequencing was assessed. Using primers within the set of the seven MLST officially adopted for identifying Xf at type strain level, we developed a workflow consisting in a multiple PCR and an ad hoc pipeline to generate MLST consensus after Nanopore-sequencing of the amplicons. The here-developed combined approach achieved a sensitivity higher than real-time PCR allowing within few hours, the detection and identification of Xf at ST level in infected plant material, also at low level of contamination.Originality Significance StatementIn this work we developed a methodology that allows the detection and identification of Xylella fastidiosa in plant using the Nanopore technology portable device MinION. The approach that we develop resulted more sensitive than methods currently used for detecting X. fastidiosa, like real-time PCR. This approach can be extensively used for X. fastidiosa detection and it may pave the road for the detection of other tedious vascular pathogens.

scholarly journals Development of Real-Time and Conventional PCR Assays for Identifying Stubby Root Nematode Paratrichodorus allius

Plant Disease ◽  
2017 ◽  
Vol 101 (6) ◽  
pp. 964-972 ◽  
Author(s):  
Danqiong Huang ◽  
Guiping Yan ◽  
Andrea M. Skantar
Keyword(s):  
Real Time ◽  
Molecular Diagnosis ◽  
Real Time Pcr ◽  
Tobacco Rattle Virus ◽  
Nematode Species ◽  
Detection Sensitivity ◽  
Conventional Pcr ◽  
Nematode Communities ◽  
Detection And Identification ◽  
Pcr Assays

Paratrichodorus allius is an important pest on many crops, particularly on potato due to its ability to transmit Tobacco rattle virus causing corky ringspot disease on tubers. Detection and identification of P. allius are important for effective disease management. In this study, a rapid and reliable molecular diagnosis of this nematode targeting internal transcribed spacer ribosomal DNA was established. The specificity of the designed primers was evaluated using 29 nematode species and results showed that a single amplicon was produced from DNA of P. allius only. Detection sensitivity analysis indicated that a 9.6 × 10−4 ng of DNA template could be detected by conventional PCR and 1.92 × 10−4 ng of DNA by real-time PCR. The PCR assays amplified DNA of stubby root nematodes isolated from 18 soil samples in North Dakota and Minnesota, which were confirmed as P. allius by sequencing. Both conventional PCR and real-time PCR assays amplified target nematodes from complex nematode communities, supporting the success of this molecular diagnosis of P. allius. This is the first report of P. allius identification using the real-time PCR method and from nematode communities with other nematodes using conventional PCR. The new PCR assays provide rapid species identification and are suitable for use in diagnostic laboratories and detection of field infestations with P. allius.

scholarly journals Designing Two Synthetic Constructs for Real Time PCR Detection of Francisella tularensis and Ebola Virus

2021 ◽  
Author(s):  
Mohammadjavad Dehghan Esmat Abadi ◽  
Hesam Motalebzadeh ◽  
Mahmoud Barati ◽  
Mohammadali Yaghobi
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Francisella Tularensis ◽  
Real Time Pcr ◽  
Ebola Virus ◽  
Positive Control ◽  
Molecular Methods ◽  
Dual Band ◽  
Conventional Pcr ◽  
Micro Organisms

Background: Generally, timely diagnosis of micro-organisms is very important to prevent many diseases. Many methods can detect micro-organisms like culture-based methods and molecular methods. The molecular methods are usually preferred because they provide fast and reliable results. In some cases, microbial strains are not accessible, and there is no safety to work with them; therefore, synthetic constructs which are designed according to the available sequences in databases can be used as a positive control for detection of them. Methods: In this study, a synthetic construct was designed for molecular detection of Francisella tularensis (F. tularensis) and the Ebola virus by multiplex real-time PCR reaction. For this, sequences were taken from databases and then multiple alignments were done by software. Also, conventional PCR and two models of real-time PCR (SYBR green and TaqMan) were applied. Finally, multiplex real-time PCR was performed. Results: The synthetic construct was designed and used for conventional PCR and multiplex PCR. The results of common PCR showed a single band at 148 bp and 167 bp in 1.5% agarose gel stained by ethidium bromide for F. tularensis and Ebola virus, respectively. Also, a dual-band at 148 and 167 bp was observed in multiplex PCR. Results of real-time PCR showed a limit of detection about 0.1 pg of plasmid/µl. Conclusion: In conclusion, the designed construct can be used as a positive control for an accurate diagnosis of these micro-organisms without any biological danger for laboratory staff. So, this method is useful for diagnosis of these agents in food, water, and blood samples.

scholarly journals The case for tracking misinformation the way we track disease

2021 ◽  
Vol 8 (1) ◽  
pp. 205395172110138
Author(s):  
Erika Bonnevie ◽  
Jennifer Sittig ◽  
Joe Smyser
Keyword(s):  
Public Health ◽  
Real Time ◽  
Supervised Machine Learning ◽  
Disease Spread ◽  
The Public ◽  
Monitoring Tools ◽  
Core Function ◽  
Public Health Organizations ◽  
Front Line Workers ◽  
Media Data

While public health organizations can detect disease spread, few can monitor and respond to real-time misinformation. Misinformation risks the public’s health, the credibility of institutions, and the safety of experts and front-line workers. Big Data, and specifically publicly available media data, can play a significant role in understanding and responding to misinformation. The Public Good Projects uses supervised machine learning to aggregate and code millions of conversations relating to vaccines and the COVID-19 pandemic broadly, in real-time. Public health researchers supervise this process daily, and provide insights to practitioners across a range of disciplines. Through this work, we have gleaned three lessons to address misinformation. (1) Sources of vaccine misinformation are known; there is a need to operationalize learnings and engage the pro-vaccination majority in debunking vaccine-related misinformation. (2) Existing systems can identify and track threats against health experts and institutions, which have been subject to unprecedented harassment. This supports their safety and helps prevent the further erosion of trust in public institutions. (3) Responses to misinformation should draw from cross-sector crisis management best practices and address coordination gaps. Real-time monitoring and addressing misinformation should be a core function of public health, and public health should be a core use case for data scientists developing monitoring tools. The tools to accomplish these tasks are available; it remains up to us to prioritize them.

scholarly journals A single-run, real-time PCR for detection and identification ofBorrelia burgdorferi sensu latospecies, based on thehbbgene sequence

2006 ◽  
Vol 259 (1) ◽  
pp. 35-40 ◽  
Author(s):  
Denis Portnoï ◽  
Natacha Sertour ◽  
Elisabeth Ferquel ◽  
Martine Garnier ◽  
Guy Baranton ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Detection And Identification

scholarly journals Snus and snus-like nicotine products moving across Nordic borders: Can laws protect young people?

2021 ◽  
pp. 145507252199570
Author(s):  
Marjut Salokannel ◽  
Eeva Ollila
Keyword(s):  
Public Health ◽  
Young People ◽  
Health Impact ◽  
The Public ◽  
European Court ◽  
Wide Availability ◽  
Personal Use ◽  
Health Studies ◽  
High Level ◽  
The Eu

Background: Use of snus and snus-like nicotine products is increasing, in particular among young people, in several Nordic countries and Estonia, while snus is legally on the market only in Sweden and Norway. Snus is available in a great variety of tastes and packaging particularly catering for young users. Recently, strong snus-resembling nicotine pouches have emerged on the market. This research investigates the regulatory means to counteract this development. Methods: European Union (EU) and national tobacco control legislation, case law of the European Court of Justice (CJEU) and relevant public health studies are analysed. Results: The research finds that the judgement of the CJEU relating to the sale of snus on Finnish ferries has not been enforced. Permitted large traveller imports for personal use have contributed to wide availability of snus in Finland. Even if the legislation in Sweden is in conformity with the exemption it obtained in the Accession Treaty, the public health impact of snus use for young people in its neighbouring countries has become considerable. Nicotine pouches, -which are not regarded as medical products in terms of medicine legislation, lack harmonised EU-wide regulation. Controlling smuggling across open borders is challenging. Conclusions: The legislation at the EU and national levels should be able to protect young people from new tobacco and nicotine products. It is urgent to harmonise regulation relating to new tobacco and nicotine products taking as a base a high level of protection of health as required in the Treaty on the Functioning of the EU.

Sign in / Sign up

Export Citation Format

Share Document