Comparison of flavivirus universal primer pairs and development of a rapid, highly sensitive heminested reverse transcription-PCR assay for detection of flaviviruses targeted to a conserved region of the NS5 gene sequences

Journal of Clinical Microbiology
N ScaramozzinoD Garin

Abstract

Arthropod-transmitted flaviviruses are responsible for considerable morbidity and mortality, causing severe encephalitic, hemorrhagic, and febrile illnesses in humans. Because there are no specific clinical symptoms for infection by a determined virus and because different arboviruses could be present in the same area, a genus diagnosis by PCR would be a useful first-line diagnostic method. The six published Flavivirus genus primer pairs localized in the NS1, NS3, NS5, and 3' NC regions were evaluated in terms of specificity and sensitivity with flaviviruses (including the main viruses pathogenic for humans) at a titer of 10(5) 50% tissue culture infectious doses (TCID(50)s) ml(-1) with a common identification step by agarose gel electrophoresis. Only one NS5 primer pair allowed the detection of all tested flaviviruses with the sensitivity limit of 10(5) TCID(50)s ml(-1). Using a heminested PCR with new primers designed in the same region after an alignment of 30 different flaviviruses, the sensitivity of reverse transcription-PCR was improved and allowed the detection of about 200 infectious doses ml(-1) with all of the tick- and mosquito-borne flaviviruses tested. It was confirmed that the sequenced amplified products in the ...Continue Reading

References

Apr 11, 1991·Journal of Medical Virology·Z A EldadahD C Gajdusek
Jan 1, 1990·Annual Review of Microbiology·T J ChambersC M Rice
Mar 22, 1985·Science·D J Lipman, W R Pearson
Mar 1, 1993·Journal of Virological Methods·M Tanaka
Oct 1, 1993·Journal of Virological Methods·L FulopR W Titball
Aug 1, 1995·Journal of Virological Methods·B H RuizS Liu
Jan 23, 1996·Proceedings of the National Academy of Sciences of the United States of America·P M ZanottoE C Holmes
Jan 7, 1998·Journal of Virology·G KunoC B Cropp
Jul 22, 1998·Journal of Virological Methods·G Kuno
Feb 19, 2000·MMWR. Morbidity and Mortality Weekly Report·UNKNOWN Centers for Disease Control and Prevention (CDC)

❮ Previous
Next ❯

Citations

May 6, 2008·Transactions of the Royal Society of Tropical Medicine and Hygiene·A P G AlmeidaA Esteves
Mar 5, 2003·Antiviral Research·Christian DrostenStephan Günther
Feb 19, 2003·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Paola CinqueAke Lundkvist
Jul 31, 2010·Epidemiology and Infection·L BusaniS Marangon
Apr 23, 2004·The New England Journal of Medicine·Poh Lian LimYee-Sin Leo
May 15, 2009·The New England Journal of Medicine·Norma P TavakoliPhyllis L Faust
Jun 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Sonja Hall-MendelinAndrew F van den Hurk
May 3, 2006·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Christopher D PaddockSherif R Zaki
Jan 19, 2006·Vector Borne and Zoonotic Diseases·Aida EstevesM Teresa Novo
Nov 21, 2007·Vector Borne and Zoonotic Diseases·G MoureauX de Lamballerie
Apr 8, 2010·Vector Borne and Zoonotic Diseases·Mattia CalzolariMichele Dottori
Sep 22, 2010·Vector Borne and Zoonotic Diseases·Nicholas JohnsonAnthony R Fooks
Sep 22, 2010·Vector Borne and Zoonotic Diseases·António Paulo G AlmeidaAida Esteves
Sep 21, 2010·Vector Borne and Zoonotic Diseases·Marco TambaPaola Angelini
Nov 21, 2012·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Marc Y El KhouryGary P Wormser
Jul 4, 2013·The Journal of Infectious Diseases·Luisa BarzonGiorgio Palù
Oct 19, 2004·Clinical Microbiology Reviews·Roberta L Debiasi, Kenneth L Tyler
Feb 8, 2005·Journal of Clinical Microbiology·Roberta Vieira de Morais BronzoniLuiz Tadeu Moraes Figueiredo
Aug 6, 2005·Journal of Clinical Microbiology·Jehanara KorimbocusGuy Vernet
Dec 8, 2009·Annual Review of Entomology·Gregory D Ebel
Mar 1, 2008·Virology Journal·Erika LindhOlli Vapalahti
Oct 20, 2010·PLoS Neglected Tropical Diseases·Cristina DomingoOliver Donoso-Mantke
Jul 23, 2013·PLoS Neglected Tropical Diseases·John A CrumpJohn A Bartlett
Jun 3, 2010·The Open Virology Journal·Marina MoniniFranco Maria Ruggeri
Sep 2, 2008·Emerging Infectious Diseases·Miguel Angel Jiménez-ClaveroUrsula Höfle
May 3, 2011·Emerging Infectious Diseases·Paolo RavaniniOlli Vapalahti
Aug 2, 2011·Emerging Infectious Diseases·Marietjie VenterFrank Freeman
Aug 2, 2011·Emerging Infectious Diseases·Montserrat AgüeroMiguel Angel Jiménez-Clavero
Jun 6, 2013·Emerging Infectious Diseases·Francisco LlorenteMiguel Angel Jiménez-Clavero
Sep 26, 2013·Viruses·Luisa BarzonGiorgio Palù

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.