Loop-Mediated Isothermal Amplification (LAMP)
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Spotlight Briefing Note
Spotlight briefing Responding to the COVID-19 pandemic Ten questions on the next phase of the UK’s COVID-19 response October 2020 Overview • There are a number of questions which remain as to the next phase of the response to the COVID-19 pandemic. • What values have informed the most recent decisions on COVID-19 restrictions? Public health measures involve a number of challenging trade-offs between different rights and interests. Alongside the scientific evidence, it must be made clear which values are guiding decisions about which, and whose, interests take priority, and why. • Is the government considering the use of “immunity certificates” in the next phase of the response? Any approach which relies on a system of ‘immunity certification’ raises a number of ethical questions concerning individual rights versus the public interest, and social justice. If the Government is considering such a system, there must be a robust and open debate now. • How will development of an effective COVID-19 vaccine affect uptake - and what should be done? Issues around the speed of development, changes in regulation, and communication with the public may all affect public trust and uptake of any vaccine. Consideration about how to address these issues should take place now. • What discussions are taking place on setting priorities for vaccine allocation within the UK? There is a range of different values which can be taken into consideration when setting priorities for access to limited doses of a vaccine. What values and interests will guide decision-making in this area must be clearly set out. • How will the UK ensure a sustained commitment to global solidarity? The global nature of the pandemic shows the importance of working as part of a global effort. -
SARS-Cov-2 RNA, Qualitative Real-Time RT-PCR (Test Code 39433)
SARS-CoV-2 RNA, Qualitative Real-Time RT-PCR (Test Code 39433) Package Insert For Emergency Use Only For In-vitro Diagnostic Use - Rx Only Intended Use The Quest Diagnostics SARS-CoV-2 RNA, Qualitative Real-Time RT-PCR (“Quest SARS-CoV-2 rRT-PCR”) is a real-time RT-PCR test intended for the qualitative detection of nucleic acid from the SARS-CoV-2 in upper and lower respiratory specimens (such as nasopharyngeal or oropharyngeal swabs, sputum, tracheal aspirates, and bronchoalveolar lavage) collected from individuals suspected of COVID-19 by their healthcare provider. This test is also for use with nasal swab specimens that are self-collected at home or in a healthcare setting by individuals using an authorized home-collection kit when determined to be appropriate by a healthcare provider. This test is for the qualitative detection of nucleic acid from the SARS-CoV-2 in pooled samples containing up to four of the individual upper respiratory swab specimens (nasopharyngeal, mid-turbinate, anterior nares or oropharyngeal swabs) that were collected in individual vials containing transport media from individuals suspected of COVID-19 by their healthcare provider. Negative results from pooled testing should not be treated as definitive. If patient’s clinical signs and symptoms are inconsistent with a negative result or results are necessary for patient management, then the patient should be considered for individual testing. Specimens included in pools with a positive, inconclusive, or invalid result must be tested individually prior to reporting a result. Specimens with low viral loads may not be detected in sample pools due to the decreased sensitivity of pooled testing. -
Whole Day Download the Hansard
Monday Volume 681 28 September 2020 No. 109 HOUSE OF COMMONS OFFICIAL REPORT PARLIAMENTARY DEBATES (HANSARD) Monday 28 September 2020 © Parliamentary Copyright House of Commons 2020 This publication may be reproduced under the terms of the Open Parliament licence, which is published at www.parliament.uk/site-information/copyright/. HER MAJESTY’S GOVERNMENT MEMBERS OF THE CABINET (FORMED BY THE RT HON. BORIS JOHNSON, MP, DECEMBER 2019) PRIME MINISTER,FIRST LORD OF THE TREASURY,MINISTER FOR THE CIVIL SERVICE AND MINISTER FOR THE UNION— The Rt Hon. Boris Johnson, MP CHANCELLOR OF THE EXCHEQUER—The Rt Hon. Rishi Sunak, MP SECRETARY OF STATE FOR FOREIGN,COMMONWEALTH AND DEVELOPMENT AFFAIRS AND FIRST SECRETARY OF STATE— The Rt Hon. Dominic Raab, MP SECRETARY OF STATE FOR THE HOME DEPARTMENT—The Rt Hon. Priti Patel, MP CHANCELLOR OF THE DUCHY OF LANCASTER AND MINISTER FOR THE CABINET OFFICE—The Rt Hon. Michael Gove, MP LORD CHANCELLOR AND SECRETARY OF STATE FOR JUSTICE—The Rt Hon. Robert Buckland, QC, MP SECRETARY OF STATE FOR DEFENCE—The Rt Hon. Ben Wallace, MP SECRETARY OF STATE FOR HEALTH AND SOCIAL CARE—The Rt Hon. Matt Hancock, MP SECRETARY OF STATE FOR BUSINESS,ENERGY AND INDUSTRIAL STRATEGY—The Rt Hon. Alok Sharma, MP SECRETARY OF STATE FOR INTERNATIONAL TRADE AND PRESIDENT OF THE BOARD OF TRADE, AND MINISTER FOR WOMEN AND EQUALITIES—The Rt Hon. Elizabeth Truss, MP SECRETARY OF STATE FOR WORK AND PENSIONS—The Rt Hon. Dr Thérèse Coffey, MP SECRETARY OF STATE FOR EDUCATION—The Rt Hon. Gavin Williamson CBE, MP SECRETARY OF STATE FOR ENVIRONMENT,FOOD AND RURAL AFFAIRS—The Rt Hon. -
Letter Tony Nikolic Wrote to New South Wales Minister
4. I am instructed to send this letter to sitting Members of Parliament (State and Federal). 5. The most recent Orders from the NSW State Government on Airport workers at Kingsford Smith Airport (Sydney) has caused a number of airport employees (including pregnant and single wage families/adults) to go on stress leave because they have been given an ultimatum to get the vaccine or potentially lose their positions. The amount of emails and calls from airline industry staff to my office is most concerning and I am instructed to make this representation on their behalf because they believe their elected representatives are not taking notice. 6. We are also mindful that the airport staff, front-line workers, pilots, police, paramedics and medical staff are also seeking the protection of all rights and responsibilities pursuant to Australian and International laws. 7. There are people taking sick leave from airport duties due to the mental health related issues and pressures being placed on them to take a vaccine. We understand that that the Australian Government cannot guarantee the vaccines safety in the short, mid and long term, but yet the NSW Government are issuing Orders for companies to vaccinate all staff, whilst Federal Parliamentarians do not question the validity or acknowledge this growing concern in electorates all over Australia. 8. I am instructed those workers are not allowed on Commonwealth airport property effective as of 6 July 2021 as a result of State Orders. The workers are not only feeling the pressures imposed upon them with speed and stealth at which the Orders were passed, but they are also concerned about being targeted if they speak out. -
Optimizing Primerá/Probe Design for Fluorescent
Journal of Neuroscience Methods 123 (2003) 31Á/45 www.elsevier.com/locate/jneumeth Optimizing primerÁ/probe design for fluorescent PCR Dmitri Proudnikov *, Vadim Yuferov, Yan Zhou, K.Steven LaForge, Ann Ho, Mary Jeanne Kreek Laboratory of the Biology of Addictive Diseases, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA Received 30 July 2002; received in revised form 21 October 2002; accepted 25 October 2002 Abstract TaqMan†,avariation of fluorescent PCR, is a powerful tool for gene expression and polymorphism studies. Here we describe the design and evaluation of 27 new TaqMan† primer-probe sets for rat genes that play a key role in neural signaling. These newly designed and synthesized probes were tested and then used for quantification of RNA isolated from rat brain. The usual length of † common TaqMan probes is 25 bases or less. In these studies we constructed probes with lengths of 25Á/39 bases to span exonÁ/exon junctions of nucleic acids to avoid the influence of DNA contamination upon the RNA quantification. The specific sequences at these positions required probes of these lengths to optimize hybridization. We found that the relocation of the quencher from the traditional 3? position to an internal one increases the sensitivity of probe up to 30 fold. Substitution of 6-carboxyfluorescein with Alexa Fluor† 488 as fluorophore and TAMRA with non-fluorescent quencher dabcyl was also investigated. We also describe the evaluation of part of a newly designed set of 27 TaqMan† primer-probes for the measurement of differences in gene expression levels in samples from the caudate putamen region of rat brain after ‘binge’ paradigm cocaine administration. -
BMJ in the News Is a Weekly Digest of Journal Stories, Plus Any Other News
BMJ in the News is a weekly digest of journal stories, plus any other news about the company that has appeared in the national and a selection of English-speaking international media. A total of 25 journals were picked up in the media last week (21-27 September) - our highlights include: ● Research in The BMJ on excess belly fat and risk of early death was covered widely, including CNN, The Daily Mail and The Daily Telegraph. ● The New York Times ran an op-ed by Peter Doshi, Associate Editor at The BMJ on covid-19 vaccine trials. ● Research in Thorax warning that patients with no symptoms carry as much covid-19 virus as those with symptoms generated global headlines, including NBC News, New York Daily News and The Guardian. BMJ PRESS RELEASES The BMJ | BMJ Open Open Heart | Thorax EXTERNAL PRESS RELEASES BMJ Open | BMJ Open Diabetes Research & Care British Journal of Sports Medicine | Emergency Medicine Journal OTHER COVERAGE The BMJ | Annals of the Rheumatic Diseases Archives of Disease in Childhood | BMJ Evidence-Based Medicine BMJ Global Health | BMJ Nutrition, Prevention & Health BMJ Open Respiratory Research | Frontline Gastroenterology Gut | Heart Injury Prevention | Journal of Epidemiology & Community Health Journal of Medical Ethics | Journal of Neurology, Neurosurgery & Psychiatry Medical Humanities | Occupational & Environmental Medicine Postgraduate Medical Journal | Practical Neurology Vet Record B MJ New Co-Editors-in-Chief for BMJ Quality -
WHO Backs Rollout of Astrazeneca Vaccine
this week TWINS page 423 • VACCINATING CHILDREN page 424 • BRITISH CYCLING page 427 FRANK HOERMANN/DPA/PA/ALAMY FRANK WHO backs rollout of AstraZeneca vaccine Doctors have warned of the risks associated On 16 March WHO’s chief scientist, Soumya The World Health Organization with pausing or delaying vaccination Swaminathan, said, “We do not want has urged people not to panic programmes against covid-19, as the people to panic, and we would, for the time amid reports of blood clotting number of European countries that have being, recommend that countries continue disorders in patients receiving the vaccine halted use of the Oxford University and vaccinating with AstraZeneca . So far, we AstraZeneca vaccine rose to 16. do not fi nd an association between these Denmark, Norway, Bulgaria, Iceland, events and the vaccine.” France, Germany, Italy, Spain, Portugal, The EMA said there had been 30 reports Slovenia, and Cyprus have suspended all of thromboembolic events among nearly use of the vaccine. Five other countries fi ve million people given the AstraZeneca (Austria, Estonia, Latvia, Lithuania, and vaccine in the European Economic Area. Luxembourg) have paused the use of a AstraZeneca has said 37 blood clots have LATEST ONLINE batch of a million doses of the vaccine. been reported among more than 17 million NHS and social The moves came after reports of people vaccinated in the EU and Britain. care need an extra blood clotting disorders. The Norwegian Five of the cases were deep vein thrombosis, £12bn to get back Medicines Agency said last week it was and 22 were pulmonary embolisms. -
Rnase-H-Based Assays Utilizing Modified Rna
(19) TZZ ¥_T (11) EP 2 279 263 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12Q 1/68 (2006.01) 04.09.2013 Bulletin 2013/36 (86) International application number: (21) Application number: 09739895.2 PCT/US2009/042454 (22) Date of filing: 30.04.2009 (87) International publication number: WO 2009/135093 (05.11.2009 Gazette 2009/45) (54) RNASE-H-BASED ASSAYS UTILIZING MODIFIED RNA MONOMERS TESTS AUF RNASE-H-BASIS MIT MODIFIZIERTEN RNA-MONOMEREN DOSAGES À BASE DE RNASE-H UTILISANT DES MONOMÈRES D’ARN MODIFIÉS (84) Designated Contracting States: • ROSE, Scott AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Coralville, IA 52241 (US) HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL • DOBOSY, Joseph PT RO SE SI SK TR Coralville, IA 52241 (US) (30) Priority: 30.04.2008 US 49204 P (74) Representative: Grünecker, Kinkeldey, Stockmair & Schwanhäusser (43) Date of publication of application: Leopoldstrasse 4 02.02.2011 Bulletin 2011/05 80802 München (DE) (60) Divisional application: (56) References cited: 13173388.3 EP-A- 1 367 136 WO-A-01/21813 13173389.1 WO-A-03/074724 WO-A-2004/059012 13173390.9 WO-A-2007/062495 WO-A2-2005/021776 13173391.7 WO-A2-2007/141580 US-A- 5 744 308 US-A- 5 830 664 (73) Proprietor: Integrated Dna Technologies, Inc. Coralville, IA 52241 (US) • ITAYA M ET AL: "Molecular cloning of a ribonuclease H (RNase HI) gene from an extreme (72) Inventors: thermophile Thermus thermophilus HB8: a • WALDER, Joseph, Alan thermostable RNase H can functionally replace Chicago, IL 60645 (US) the Escherichia coli enzyme in vivo." NUCLEIC • BEHLKE, Mark, Aaron ACIDS RESEARCH 25 AUG 1991, vol. -
Essentials of Real Time PCR About Sequence Detection Chemistries
Essentials of Real Time PCR About Real-Time PCR Assays Real-time Polymerase Chain Reaction (PCR) is the ability to monitor the progress of the PCR as it occurs ( i.e., in real time). Data is therefore collected throughout the PCR process, rather than at the end of the PCR. This completely revolutionizes the way one approaches PCR-based quantitation of DNA and RNA. In real-time PCR, reactions are characterized by the point in time during cycling when amplification of a target is first detected rather than the amount of target accumulated after a fixed number of cycles. The higher the starting copy number of the nucleic acid target, the sooner a significant increase in fluorescence is observed. In contrast, an endpoint assay (also called a “plate read assay”) measures the amount of accumulated PCR product at the end of the PCR cycle. About Sequence Detection Chemistries Overview Applied Biosystems has developed two types of chemistries used to detect PCR products using Sequence Detection Systems (SDS) instruments: ® • TaqMan chemistry (also known as “fluorogenic 5´ nuclease chemistry”) ® • SYBR Green I dye chemistry TaqMan® Chemistry The TaqMan chemistry uses a fluorogenic probe to enable the detection of a specific PCR product as it accumulates during PCR cycles. Assay Types that Use TaqMan Chemistry The TaqMan chemistry can be used for the following assay types: • Quantitation, including: – One-step RT-PCR for RNA quantitation – Two-step RT-PCR for RNA quantitation – DNA/cDNA quantitation • Allelic Discrimination • Plus/Minus using an IPC SYBR Green I Dye Chemistry The SYBR Green I dye chemistry uses SYBR Green I dye, a highly specific, double-stranded DNA binding dye, to detect PCR product as it accumulates during PCR cycles. -
Operation Moonshot Proposals Are Scientifically Unsound
EDITORIALS BMJ: first published as 10.1136/bmj.m3699 on 22 September 2020. Downloaded from 1 University of Birmingham, Operation Moonshot proposals are scientifically unsound Birmingham, UK They could do more harm than good to people, populations, and the economy 2 University of Leicester. Leicester, UK Jonathan J Deeks, 1 Anthony J Brookes, 2 Allyson M Pollock3 3 Newcastle University, Newcastle, UK Correspondence to: J Deeks The polymerase chain reaction (PCR) swab test is balance of costs and harms against the potential [email protected] useful (but not perfect) for detecting SARS-CoV-2 benefits has not been evaluated Cite this as: BMJ 2020;370:m3699 virus RNA in symptomatic patients.1 However, Testing conundrums http://dx.doi.org/10.1136/bmj.m3699 problems arise using the test for purposes that Published: 22 September 2020 disregard symptoms or time of infection—namely, Now, Operation Moonshot has proposed that mass case finding, mass screening, and disease screening with less accurate point-of-care tests will surveillance. help “reduce the ‘R’ rate, keep the economy open and enable a return to normal life.”10 Could this work? This is because PCR is not a test of infectiousness. Rather, the test detects trace amounts of viral genome The Moonshot proposals are based exclusively on sequence, which may be either live transmissible computer modelling11 not empirical evidence. virus or irrelevant RNA fragments from previous Critically, the model considers repeated use of tests infection.2 When people with symptoms or who have that are positive only in infected people with high been recently exposed receive a positive PCR result viral loads of SARS-CoV-2. -
The Time the Children Didn't Go to School
THE TIME THE CHILDREN DIDN’T GO TO SCHOOL ANNABELLE HAYES FOREWORD ......................................................... 3 ACKNOWLEDGEMENTS .................................. 4 APRIL 2020 ............................................................ 5 MAY, 2020 ............................................................ 33 JUNE, 2020 .......................................................... 63 JULY, 2020 ......................................................... 102 AUGUST, 2020 .................................................... 110 SEPTEMBER, 2020 ............................................ 114 OCTOBER, 2020 ............................................... 129 NOVEMBER, 2020 ........................................... 152 DECEMBER, 2020 ............................................ 166 JANUARY, 2021 ................................................. 176 FEBRUARY, 2021 .............................................. 202 MARCH, 2021 .................................................... 223 AFTERWORD ................................................... 230 2 FOREWORD In March 2020, schools, nurseries and colleges in the United Kingdom were shut down in response to the ongoing coronavirus pandemic. By 20 March, all schools in the UK had closed to all children except those of key workers and children considered vulnerable. After a month of numbness at having all the children home, I started these diaries to document the unprecedented time when the children didn’t go to school. When the world stopped, the children didn’t – this records their -
Use of Vectorette and Subvectorette PCR to Isolate Transgene Flanking DNA
Downloaded from genome.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press Use of Vectorette and Subvectorette PCR to Isolate Transgene Flanking DNA Maxine J. Allen, Andrew Collick, and Alec J. Jeffreys Department of Genetics, University of Leicester, Leicester LE1 7RH, UK Vectorette PCR permits the specific PCR allows specific amplification of structures and in the recovery of un- amplification of DNA segments flank- DNA fragments between two regions of known bacterial sequences. ~4-6~ In this ing a known DNA sequence. It en- known sequence to which primers can paper we detail its use in the systematic ables the application of the PCR be directed. Often, however, molecular isolation of 5'- and 3'-flanking junction where sequence information is only biological techniques require the isola- sequences around mouse transgenes of available for one primer site. We now tion and subsequent sequence analysis the minisatellite MS32. MS32 is a hu- show that vectorette PCR can be used of unknown regions of DNA. PCR can be man minisatellite composed of a tan- for the systematic mapping and re- used to retrieve this unknown sequence dem array of 29-bp repeat-unit sequence trieval of transgene flanking DNA. if it is flanking a region of known se- variants. The interspersion of these We also show that the sequence of quence. In this case, a primer can be de- sequence variants can be assayed in large vectorette PCR fragments can signed in either orientation from the the minisatellite variant repeat (MVR)- be obtained without cloning, by the known sequence.