scholarly journals Development and application of a real-time RT-PCR assay to rapidly detect H2 subtype avian influenza A viruses

2021 ◽  
pp. 104063872199481
Author(s):  
Yixin Xiao ◽  
Fan Yang ◽  
Fumin Liu ◽  
Hangping Yao ◽  
Nanping Wu ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A ◽  
Minor Groove Binder ◽  
Rt Pcr ◽  
Influenza A Viruses ◽  
Pcr Assay ◽  
Infectious Dose ◽  
The Matrix ◽  
Taqman Minor Groove Binder

The H2 subtypes of avian influenza A viruses (avian IAVs) have been circulating in poultry, and they have the potential to infect humans. Therefore, establishing a method to quickly detect this subtype is pivotal. We developed a TaqMan minor groove binder real-time RT-PCR assay that involved probes and primers based on conserved sequences of the matrix and hemagglutinin genes. The detection limit of this assay was as low as one 50% egg infectious dose (EID50)/mL per reaction. This assay is specific, sensitive, and rapid for detecting avian IAV H2 subtypes.

scholarly journals Development and evaluation of a one-step real-time RT-PCR assay for universal detection of influenza A viruses from avian and mammal species

2010 ◽  
Vol 155 (5) ◽  
pp. 665-673 ◽  
Author(s):  
Alexander Nagy ◽  
Veronika Vostinakova ◽  
Zuzana Pirchanova ◽  
Lenka Cernikova ◽  
Zuzana Dirbakova ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza A ◽  
Rt Pcr ◽  
Influenza A Viruses ◽  
Pcr Assay ◽  
Mammal Species ◽  
One Step ◽  
Universal Detection

scholarly journals Development and Evaluation of a SYBR Green-Based Real Time RT-PCR Assay for Detection of the Emerging Avian Influenza A (H7N9) Virus

PLoS ONE ◽  
2013 ◽  
Vol 8 (11) ◽  
pp. e80028 ◽  
Author(s):  
Zheng Zhu ◽  
Huan Fan ◽  
Xian Qi ◽  
Yuhua Qi ◽  
Zhiyang Shi ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A ◽  
Sybr Green ◽  
H7n9 Virus ◽  
Rt Pcr ◽  
Pcr Assay

Development and evaluation of a real-time RT-PCR assay for detection of a novel avian influenza A (H5N6) virus

2018 ◽  
Vol 257 ◽  
pp. 79-84 ◽  
Author(s):  
Rusheng Zhang ◽  
Dong Yao ◽  
Jingfang Chen ◽  
Wen Ye ◽  
Xinhua Ou ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A ◽  
Rt Pcr ◽  
Pcr Assay

scholarly journals Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one-step real-time RT-PCR assay

2014 ◽  
Vol 14 (1) ◽  
Author(s):  
Jian Fan ◽  
David Cui ◽  
Siuying Lau ◽  
Guoliang Xie ◽  
Xichao Guo ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A ◽  
H7n9 Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

Real-time RT-PCR assay to differentiate clades of H5N1 avian influenza viruses circulating in Vietnam

2013 ◽  
Vol 193 (2) ◽  
pp. 452-458 ◽  
Author(s):  
Z. Kis ◽  
J. Jones ◽  
A. Creanga ◽  
K. Ferdinand ◽  
K. Inui ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Avian Influenza Viruses ◽  
Pcr Assay ◽  
H5n1 Avian Influenza

scholarly journals Development of a Diagnostic System for Novel Influenza A(H7N9) Virus Using a Real-Time RT-PCR Assay in Japan

2015 ◽  
Vol 68 (2) ◽  
pp. 113-118 ◽  
Author(s):  
Ikuyo Takayama ◽  
Hitoshi Takahashi ◽  
Mina Nakauchi ◽  
Shiho Nagata ◽  
Masato Tashiro ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza A ◽  
Diagnostic System ◽  
H7n9 Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Novel Influenza A

scholarly journals Real-Time Reverse Transcription-Polymerase Chain Reaction Detection of Newcastle Disease Virus Using Light upon Extension Fluorogenic Primers

2007 ◽  
Vol 19 (4) ◽  
pp. 400-404 ◽  
Author(s):  
Márta Antal ◽  
Tibor Farkas ◽  
Péter Germán ◽  
Sándor Belák ◽  
István Kiss
Keyword(s):  
Newcastle Disease Virus ◽  
Real Time ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Plasmid Copy Number ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Plasmid Copy ◽  
Infectious Dose ◽  
Efficient Detection

A real-time reverse transcriptase (RT)-PCR assay, applying light upon extension (LUX) fluorogenic primers, was developed for rapid and efficient detection of Newcastle disease virus (NDV). The method, which targets the fusion (F) protein gene of the viral genome, gave positive signal with all NDV isolates tested (32/32), while negative results were obtained with heterologous pathogens (35/35), including 13 avian influenza virus isolates. The detection limit of the assay was approximately 10+1.2 egg infectious dose (EID)50/0.2 ml and 10+2.2 EID50/0.2 ml for virus suspensions and spiked chicken fecal samples, respectively. As expressed in plasmid copy number, the procedure has a sensitivity of approximately 20 copies of the plasmid harboring the target gene. Due to its high specificity, sensitivity, and relative simplicity, the LUX RT-PCR assay provides a novel, rapid, and practical tool for the detection of NDV.

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.

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