Specific detection of the novel goose astrovirus using a TaqMan real-time RT-PCR technology

2019 ◽  
Vol 137 ◽  
pp. 103766 ◽  
Author(s):  
Chunhe Wan ◽  
Cuiteng Chen ◽  
Longfei Cheng ◽  
Guanghua Fu ◽  
Shaohua Shi ◽  
...  
Keyword(s):  
Real Time ◽  
The Novel ◽  
Rt Pcr ◽  
Specific Detection ◽  
Pcr Technology

Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression’s C T difference” formula

2006 ◽  
Vol 84 (11) ◽  
pp. 901-910 ◽  
Author(s):  
Jan H. Schefe ◽  
Kerstin E. Lehmann ◽  
Ivo R. Buschmann ◽  
Thomas Unger ◽  
Heiko Funke-Kaiser
Keyword(s):  
Data Analysis ◽  
Real Time ◽  
The Novel ◽  
Rt Pcr ◽  
Novel Gene ◽  
Difference Formula

scholarly journals Molecular Detection of a Novel Human Influenza (H1N1) of Pandemic Potential by Conventional and Real-Time Quantitative RT-PCR Assays

2009 ◽  
Vol 55 (8) ◽  
pp. 1555-1558 ◽  
Author(s):  
Leo L M Poon ◽  
K H Chan ◽  
G J Smith ◽  
C S W Leung ◽  
Y Guan ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza A ◽  
H1n1 Virus ◽  
Influenza Viruses ◽  
The Novel ◽  
Human Influenza ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Recent Emergence ◽  
Human H5n1

Abstract Background: Influenza A viruses are medically important viral pathogens that cause significant mortality and morbidity throughout the world. The recent emergence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed. Methods: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-time RT-PCR assay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RT-PCR that can discriminate the novel H1N1 from other swine and human H1 subtype viruses. Results: All of the assays had detection limits for the positive control in the range of 1.0 × 10−4 to 2.0 × 10−3 of the median tissue culture infective dose. Assay specificities were high, and for the conventional and real-time assays, all negative control samples were negative, including 7 human seasonal H1N1 viruses, 1 human H2N2 virus, 2 human seasonal H3N2 viruses, 1 human H5N1 virus, 7 avian influenza viruses (HA subtypes 4, 5, 7, 8, 9, and 10), and 48 nasopharyngeal aspirates (NPAs) from patients with noninfluenza respiratory diseases; for the assay that discriminates the novel H1N1 from other swine and human H1 subtype viruses, all negative controls were also negative, including 20 control NPAs, 2 seasonal human H1N1 viruses, 2 seasonal human H3N2 viruses, and 2 human H5N1 viruses. Conclusions: These assays appear useful for the rapid diagnosis of cases with the novel H1N1 virus, thereby allowing better pandemic preparedness.

scholarly journals Development of a Real-Time Loop-Mediated Isothermal Amplification Method for the Detection of West Nile Virus

2020 ◽  
Vol 13 (10) ◽  
Author(s):  
Jae Woong Lee ◽  
Yu-Jung Won ◽  
Sung-Geun Lee ◽  
Soon-Young Paik
Keyword(s):  
West Nile Virus ◽  
Real Time ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Genomic Sequences ◽  
The Novel ◽  
Rt Pcr ◽  
West Nile ◽  
The Real ◽  
E Gene

Background: The West Nile Virus (WNV), discovered in New York, USA in 1999 after it was first isolated in Uganda in 1937, has since spread not only in the United States but also around the world. Africa, Eurasia, Australia, and the Middle East have sporadic cases of the disease. Objectives: We aimed to find real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to be more sensitive than conventional RT-PCR, and more rapid and efficient than conventional RT-PCR and real-time RT-PCR for WNV detection. Methods: A total of 32 genomic sequences from different strains of WNV were analyzed to identify conserved nucleotide sequence regions. Six WNV specific RT-LAMP primers targeting the E gene were designed. Results: The novel primer for the real-time RT-LAMP assay can detect WNV with high specificity. The efficiency of the real-time RT-LAMP assay is higher than the conventional RT-PCR and real-time RT-PCR. Real-time RT-PCR and conventional PCR require at least 30 – 40 min and 2 h, respectively, to yield results, whereas real-time RT-LAMP provides positive results in only 10 – 20 min. Conclusions: The novel primers were developed by analyzing of 32 genomic sequences of WNV strains. The primers were designed from the most conserved region of the E gene for real-time RT-LAMP. The LAMP assay is a rapid, efficient, highly sensitive, and specific tool for the identification of WNV.

A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus

2011 ◽  
Vol 171 (2) ◽  
pp. 401-404 ◽  
Author(s):  
Carrie A. Batten ◽  
Ashley C. Banyard ◽  
Donald P. King ◽  
Mark R. Henstock ◽  
Lorraine Edwards ◽  
...  
Keyword(s):  
Real Time ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Specific Detection ◽  
Pcr Assay

Evaluation of a New Commercial Assay Using a One-Step Real Time RT-PCR Technology for HDV RNA Viral Load Quantification

2016 ◽  
Vol 64 (2) ◽  
pp. S360
Author(s):  
F. Le Gal ◽  
M. Fabrice ◽  
F. Neri Pinto ◽  
E. Anouhal ◽  
S. Brichler ◽  
...  
Keyword(s):  
Viral Load ◽  
Real Time ◽  
Rt Pcr ◽  
Commercial Assay ◽  
One Step ◽  
Pcr Technology ◽  
Viral Load Quantification

scholarly journals Laboratory preparedness in EU/EEA countries for detection of novel avian influenza A(H7N9) virus, May 2013

2014 ◽  
Vol 19 (4) ◽  
Author(s):  
E Broberg ◽  
D Pereyaslov ◽  
M Struelens ◽  
D Palm ◽  
A Meijer ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A Virus ◽  
Influenza A ◽  
Virus Detection ◽  
Influenza Viruses ◽  
World Health ◽  
Reference Laboratory ◽  
The Novel ◽  
Rt Pcr

Following human infections with novel avian influenza A(H7N9) viruses in China, the European Centre for Disease Prevention and Control, the World Health Organization (WHO) Regional Office for Europe and the European Reference Laboratory Network for Human Influenza (ERLI-Net) rapidly posted relevant information, including real-time RT-PCR protocols. An influenza RNA sequence-based computational assessment of detection capabilities for this virus was conducted in 32 national influenza reference laboratories in 29 countries, mostly WHO National Influenza Centres participating in the WHO Global Influenza Surveillance and Response System (GISRS). Twenty-seven countries considered their generic influenza A virus detection assay to be appropriate for the novel A(H7N9) viruses. Twenty-two countries reported having containment facilities suitable for its isolation and propagation. Laboratories in 27 countries had applied specific H7 real-time RT-PCR assays and 20 countries had N9 assays in place. Positive control virus RNA was provided by the WHO Collaborating Centre in London to 34 laboratories in 22 countries to allow evaluation of their assays. Performance of the generic influenza A virus detection and H7 and N9 subtyping assays was good in 24 laboratories in 19 countries. The survey showed that ERLI-Net laboratories had rapidly developed and verified good capability to detect the novel A(H7N9) influenza viruses.

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