scholarly journals Ladder-shape melting temperature isothermal amplification of nucleic acids

BioTechniques ◽  
2021 ◽  
Author(s):  
Deguo Wang ◽  
Yongzhen Wang ◽  
Meng Zhang ◽  
Yongqing Zhang ◽  
Juntao Sun ◽  
...  

A novel method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed in this study. As a proof of concept, one pair of primers or two pairs of nested primers and a thermostable DNA polymerase were employed to amplify the internal transcribed spacer of Oryza sativa with the ladder-shape melting temperature curve. Our results demonstrated that the LMTIA assay with nested primers was 50-fold more sensitive than the LAMP assay with the same level of specificity. The LMTIA method has the potential to be used for the prevention and control of emerging epidemics caused by different types of pathogens.

2020 ◽  
Author(s):  
Deguo Wang ◽  
Yongzhen Wang ◽  
Meng Zhang ◽  
Yongqing Zhang ◽  
Juntao Sun ◽  
...  

AbstractA novel method termed isothermal amplification of nucleic acids with ladder-shape melting curve (LMCP) was developed in the study. In this method, one pair of primers or two pairs of nested primers and a thermostable DNA polymerase (large fragment) were employed to amplify the internal transcribed spacer (ITS) of Oryza sativa with ladder-shape melting curve. Our results demonstrated that the LMCP assay with nested primers was 50-fold more sensitive and one-hour faster than the LAMP assay with the same level of specificity. The LMCP method has the potential to be used for the prevention and control of the emerging epidemics.


2020 ◽  
Author(s):  
Deguo Wang

AbstractBackgroundRapid and reliable diagnostic assays were critical for prevention and control of the coronavirus pneumonia caused by COVID-19.ObjectiveThis study was to establish one-pot real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay and one-pot visual RT-LAMP assay for the detection of COVID-19.MethodsSix specific LAMP primers targeting the N gene of COVID-19 were designed, the RT-LAMP reaction system was optimized with plasmid pUC57 containing N gene sequence, the detection limit was determined with a serial dilution of the plasmid pUC57 containing N gene sequence, and the one-pot real-time RT-LAMP assay and one-pot visual RT-LAMP assay for the detection of COVID-19 were established.ResultsOur results showed that the one-pot RT-LAMP assays can detect COVID-19 with a limit of ≥ 6 copies per μl−1 of pUC57 containing N gene sequence.ConclusionThis study provides rapid, reliable and sensitive tools for facilitating preliminary and cost-effective prevention and control of COVID-19.


Author(s):  
Y. Arockia Suganthi ◽  
Chitra K. ◽  
J. Magelin Mary

Dengue fever is a painful mosquito-borne infection caused by different types of virus in various localities of the world. There is no particular medicine or vaccine to treat person suffering from dengue fever. Dengue viruses are transmitted by the bite of female Aedes (Ae) mosquitoes. Dengue fever viruses are mainly transmitted by Aedes which can be active in tropical or subtropical climates. Aedes Aegypti is the key step to avoid infection transmission to save millions of people in all over the world. This paper provides a standard guideline in the planning of dengue prevention and control measures. At the same time gives the priorities including clinical management and hospitalized dengue patients have to address essentially.


Biology ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1136
Author(s):  
Caiyun Xiao ◽  
Rongyu Li

Soft rot causing Fusarium oxysporum is one of the most destructive diseases of Dendrobium officinale Kimura et Migo in China that reduces D. officinale yield and quality. A key challenge for an integrated management strategy for this disease is the rapid and accurate detection of F. oxysporum on D. officinale. Therefore, a new loop-mediated isothermal amplification (LAMP) assay was developed for this purpose. In this study, the primers were selected and designed using the translation elongation factor-1α (TEF-1α) gene region as the target DNA sequence in order to screen the best system of reaction of LAMP to detect F. oxysporum through optimizing different conditions of the LAMP reaction, including time, temperature, concentrations of MgSO4, and concentrations of inner and outer primers. The optimized system was able to efficiently amplify the target gene at 62 °C for 60 min with 1.2 μM internal primers, 0.4 μM external primers, 7 mM Mg2+, and 5 fg/µL minimum detection concentration of DNA for F. oxysporum. The amplified products could be detected with the naked eye after completion of the reaction with SYBR green I. We were better able to control the effect of soft rot in D. officinale using fungicides following a positive test result. Additionally, the control effect of synergism combinations against soft rot was higher than 75%. Thus, LAMP assays could detect F. oxysporum in infected tissues of D. officinale and soils in field, allowing for early diagnosis of the disease.


2019 ◽  
Vol 37 (2) ◽  
pp. 131-155 ◽  
Author(s):  
Willem Faes ◽  
Steven Lecompte ◽  
Zaaquib Yunus Ahmed ◽  
Johan Van Bael ◽  
Robbe Salenbien ◽  
...  

AbstractIn many industries and processes, heat exchangers are of vital importance as they are used to transfer heat from one fluid to another. These fluids can be corrosive to heat exchangers, which are usually made of metallic materials. This paper illustrates that corrosion is an important problem in the operation of heat exchangers in many environments, for which no straightforward answer exists. Corrosion failures of heat exchangers are common, and corrosion often involves high maintenance or repair costs. In this review, an overview is given of what is known on corrosion in heat exchangers. The different types of corrosion encountered in heat exchangers and the susceptible places in the devices are discussed first. This is combined with an overview of failure analyses for each type of corrosion. Next, the effect of heat transfer on corrosion and the influence of corrosion on the thermohydraulic performances are discussed. Finally, the prevention and control of corrosion is tackled. Prevention goes from general design considerations and operation guidelines to the use of cathodic and anodic protection.


Author(s):  
Lucía CASADO CASADO ◽  
Josep Ramon FUENTES I GASO

LABURPENA: Lan honetan, Europako Parlamentuaren eta Kontseiluaren 2010/75/EB Zuzentarauaren eragina aztertzen da. Horretarako, lehenik eta behin, Zuzentarauaren inguruabarrak eta alderdi azpimarragarrienak adierazten dira, eta, ondoren, ingurumen-ikuskaritzaren araubidean murgiltzen da. Oso gai berria da, ingurumen-ikuskaritzari buruzko gutxieneko araudi bat sortu den lehenengo aldia baita; orain arte, arau-baliorik gabeko Europako gomendio bat besterik ez zegoen. Hain zuzen ere, Estatu kideei gai horretan ezarritako betebeharrak aztertzen dira lanean; instalazioen gaineko ingurumen-ikuskaritzako sistema bat ezartzekoa, eta aurrezarritako ingurumen ikuskaritzako programen oinarrian egongo diren ikuskaritza planak osatzekoa, instalazioak bisitatzeko maiztasuna instalazioaren beraren motaren arabera finkatuta. Era berean, Zuzentaraua Espainiara egokitzeari buruzko oharrak egiten dira, aintzat hartuta orain dela gutxi ekainaren 11ko 5/2013 Legea onartu dela, kutsaduraren prebentzio eta kontrol integratuari buruzko uztailaren 1eko 16/2002 Legea eta hondakinen eta lurzoru kutsatuen gaineko uztailaren 28ko 22/2011 Legea aldatzen dituena. Lege berri horren eta egun izapidetzen ari diren garapen-erregelamenduen bidez, bermatuta geldituko da Zuzentarauaren egokitzea. RESUMEN: Este trabajo analiza el impacto de la Directiva 2010/75/UE del Parlamento Europeo y del Consejo, de 24 de noviembre de 2010, sobre las emisiones industriales en el régimen jurídico de la inspección ambiental. Con este fin, tras realizar una aproximación general a esta Directiva y señalar sus aspectos más destacables, se centra en el examen de la regulación de la inspección ambiental en ella contenida, aspecto muy novedoso por ser la primera vez que se incluye una regulación mínima de la inspección ambiental, que hasta ahora únicamente había sido objeto —a nivel comunitario— de una recomendación sin valor normativo, en una Directiva. Concretamente, se analizan las obligaciones establecidas para los Estados miembros en esta materia, que se traducen principalmente en la obligación de establecer un sistema de inspección medioambiental de las instalaciones y de formular planes de inspección a partir de los cuales se elaborarán programas de inspección medioambiental prefijada, que incluyan la frecuencia de las visitas a los emplazamientos para los distintos tipos de instalaciones. También se realizan algunos apuntes sobre la transposición de esta Directiva en España, teniendo en cuenta que recientemente se ha aprobado la Ley 5/2013, de 11 de junio, por la que se modifican la Ley 16/2002, de 1 de julio, de prevención y control integrados de la contaminación y la Ley 22/2011, de 28 de julio, de residuos y suelos contaminados, que, junto con su reglamento de desarrollo —actualmente en trámite—, garantizará la transposición de la Directiva. ABSTRACT: This study analyses the impact of Directive 2010/75/EU of the European Parliament and the Council, of 24 November 2010, on industrial emissions on the legislation governing environmental inspection. To this end, after providing a general discussion of the Directive and pointing out its most outstanding features, we focus on the regulation of environmental inspection that it contains. This is its most novel feature because it is the first time that a minimal regulation of environmental inspection has been included in a directive (to date, within the community, it had only been the object of a recommendation of no normative value). In particular, our study focuses on the obligations of the member states in this area, which are largely to set up a system by which installations can be environmentally inspected and inspection plans drafted on the basis of which programmes for routine environmental inspection will be drawn up that include the frequency of site visits for different types of installations. Also discussed is the transposition of the Directive in Spain, bearing in mind that Law 5/2013, of 11 June, has recently been passed to modify Law 16/2002, of 1 July, on the integrated prevention and control of pollution, and Law 22/2011, of 28 July on waste and polluted soil which, in conjunction with their regulation for implementation, guarantee the transposition of the Directive.


Author(s):  
M. A. Samad

Background: The 25.7 million cattle populations in Bangladesh which is endangered by different diseases and disorders that are associated with significant socio-economic losses to the smallholder farmers and even in modern dairy farms. More than 100 articles on the retrospective evaluation of clinical case records have been published during the last 50 years from 1968 to 2018 in Bangladesh and their compilation and evaluation would help to understand the predominant diseases associated with morbidity and mortality. Objectives: To analyze and evaluate the 50-year data on the prevalence of clinical diseases and disorders of cattle to identify the predominant and priority ailments for their prevention and control. Materials and Methods: Data obtained from all the available 118 articles published on clinical cases of cattle during the last 50 years in Bangladesh were analyzed to detect their status on bovine health. Results: Out of 137 different types of clinical cases recorded during the last 50 years, of which, calves affected with 27 (19.71%), adult cattle with 76 (55.47%) and both calves and adult cattle with 34 (24.82%) types of diseases and disorders. The highly statistical significant (p < 0001) percentage of clinical cases of cattle required medicinal treatment (81.19%) in comparison to surgical (10.03%) and gynaeco-obstetrical (GO; 08.78%) intervention. Results revealed that the parasitic diseases (fascioliasis 66.16%, humpsore 22.35%, gastro-intestinal (GI) nematodiasis 17.44%, ascariasis 12.17%, tick infestation 15.71%) and GI disorders (anorexia 14.90%, indigestion 6.15%) were highly prevalent as clinical cases in comparison to infectious diseases (anthrax 0.21%, BQ 0.92% and HS 0.45%) except FMD (10.13%) and mastitis (6.46%). Anestrus (3.04%), retained placenta (2.73%), repeat breeding (2.73%) and pyometra (2.12%) were recorded as major gynaeco-obstetrical problems, whereas umbilical myiasis (10.11%), lameness (4.25%) and abscess (1.15%) that mainly required surgical interventions. Conclusions: The analysis of 50-year published reports on clinical cases of cattle gives an overall comprehensive idea about the disease problems that would help to formulate the devices for their prevention and control. Keywords: Prevalence, Clinical diseases and disorders, 50-year review, Calf diseases, Adult cattle diseases, Medicinal cases, Gynaeco-obstetrical cases, Surgical cases


2009 ◽  
Vol 3 (3) ◽  
pp. 204-208 ◽  
Author(s):  
Alexandre Leopold Busse ◽  
Gislaine Gil ◽  
José Maria Santarém ◽  
Wilson Jacob Filho

Abstract Physical activity has been indicated as a strategy to promote health in the elderly, as well as to encourage the maintenance of functional capacity, and acts in the prevention and control of various diseases. In recent years, there has been great interest in studying the benefits of physical activity in the preservation or even improvement of cognitive performance in both the elderly without cognitive impairment and in elderly patients with some degree of cognitive impairment or dementia. The majority of epidemiological studies and clinical trials have evaluated aerobic exercises while few have assessed resistance exercise programs. The objective of this review was to examine the effects of different types of physical activity on cognitive function of elderly individuals with or without prior impairment.


2021 ◽  
Author(s):  
Ya-Ling Tan ◽  
A-Qian Huang ◽  
Li-Juan Tang ◽  
Jian-Hui Jiang

Highly sensitive digital nucleic acid techniques are of great significance for the prevention and control of epidemic diseases. Here we report the development of multiplex droplet loop-mediated isothermal amplification (multiplexed...


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