Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification

2009 ◽  
Vol 19 ◽  
Author(s):  
Man Ki Song
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Pcr Detection ◽  
Influenza Viruses ◽  
Rna Purification

scholarly journals Multiplex, Quantitative, Reverse Transcription PCR Detection of Influenza Viruses Using Droplet Microfluidic Technology

Micromachines ◽  
2014 ◽  
Vol 6 (1) ◽  
pp. 63-79 ◽  
Author(s):  
Ravi Prakash ◽  
Kanti Pabbaraju ◽  
Sallene Wong ◽  
Anita Wong ◽  
Raymond Tellier ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Pcr Detection ◽  
Influenza Viruses ◽  
Quantitative Reverse Transcription Pcr ◽  
Reverse Transcription Pcr ◽  
Microfluidic Technology

Development of a nested PCR detection method for Ranavirus

2013 ◽  
Vol 18 (3) ◽  
pp. 661-666
Author(s):  
Min ZHANG ◽  
Xiangmei LIN ◽  
Yuin JIANG
Keyword(s):  
Nested Pcr ◽  
Detection Method ◽  
Pcr Detection

Semi-Nested PCR Detection ofBartonella henselaeinIxodes persulcatusTicks from Western Siberia, Russia

2004 ◽  
Vol 4 (4) ◽  
pp. 306-309 ◽  
Author(s):  
O.V. Morozova ◽  
F.C. Cabello ◽  
A.K. Dobrotvorsky
Keyword(s):  
Nested Pcr ◽  
Western Siberia ◽  
Pcr Detection

A reverse transcription-PCR for subtyping of the neuraminidase of avian influenza viruses

2009 ◽  
Vol 155 (2) ◽  
pp. 193-198 ◽  
Author(s):  
Bao-Feng Qiu ◽  
Wu-Jie Liu ◽  
Da-Xin Peng ◽  
Shun-Lin Hu ◽  
Ying-Hua Tang ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Reverse Transcription ◽  
Influenza Viruses ◽  
Avian Influenza Viruses ◽  
Reverse Transcription Pcr

scholarly journals Multiplex Real-Time Reverse-Transcription Polymerase Chain Reaction Assays for Diagnostic Testing of Severe Acute Respiratory Syndrome Coronavirus 2 and Seasonal Influenza Viruses: A Challenge of the Phase 3 Pandemic Setting

2020 ◽  
Author(s):  
Fabiola Mancini ◽  
Fabrizio Barbanti ◽  
Maria Scaturro ◽  
Stefano Fontana ◽  
Angela Di Martino ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Real Time ◽  
Reverse Transcription ◽  
Seasonal Influenza ◽  
Influenza Viruses ◽  
Activity Period ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Pandemic coronavirus disease 2019 (COVID-19) disease represents a challenge for healthcare structures. The molecular confirmation of samples from infected individuals is crucial and therefore guides public health decision making. Clusters and possibly increased diffuse transmission could occur in the context of the next influenza season. For this reason, a diagnostic test able to discriminate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from influenza viruses is urgently needed. Methods A multiplex real-time reverse-transcription polymerase chain reaction (PCR) assay was assessed using 1 laboratory protocol with different real-time PCR instruments. Overall, 1000 clinical samples (600 from samples SARS-CoV-2–infected patients, 200 samples from influenza-infected patients, and 200 negative samples) were analyzed. Results The assay developed was able to detect and discriminate each virus target and to intercept coinfections. The limit of quantification of each assay ranged between 5 and 10 genomic copy numbers, with a cutoff value of 37.7 and 37.8 for influenza and SARS-CoV-2 viruses, respectively. Only 2 influenza coinfections were detected in COVID-19 samples. Conclusions This study suggests that multiplex assay is a rapid, valid, and accurate method for the detection of SARS-CoV-2 and influenza viruses in clinical samples. The test may be an important diagnostic tool for both diagnostic and surveillance purposes during the seasonal influenza activity period.

scholarly journals Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys

2014 ◽  
Vol 13 (1) ◽  
pp. 175 ◽  
Author(s):  
Peipei Li ◽  
Zhenjun Zhao ◽  
Ying Wang ◽  
Hua Xing ◽  
Daniel M Parker ◽  
...  
Keyword(s):  
Filter Paper ◽  
Nested Pcr ◽  
Pcr Detection ◽  
Blood Filter ◽  
Epidemiological Surveys

scholarly journals Rapid and Sensitive Detection of Noroviruses by Using TaqMan-Based One-Step Reverse Transcription-PCR Assays and Application to Naturally Contaminated Shellfish Samples

2005 ◽  
Vol 71 (4) ◽  
pp. 1870-1875 ◽  
Author(s):  
Narayanan Jothikumar ◽  
James A. Lowther ◽  
Kathleen Henshilwood ◽  
David N. Lees ◽  
Vincent R. Hill ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Specificity And Sensitivity ◽  
Routine Monitoring ◽  
The Family ◽  
One Step ◽  
Stool Specimens ◽  
Pcr Assays

ABSTRACT Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT-PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples.

HIGHLY SENSITIVE H7N9 AVIAN INFLUENZA VIRUS DETECTION USING REVERSE TRANSCRIPTION LOOP-MEDIATED ISOTHERMAL AMPLIFICATION AND OLIGONUCLEOTIDE MICROARRAY

2015 ◽  
Vol 41 (04) ◽  
pp. 251-255
Author(s):  
Lih-Chiann Wang ◽  
Dean Huang
Keyword(s):  
Avian Influenza ◽  
Sensitivity And Specificity ◽  
Reverse Transcription ◽  
High Sensitivity ◽  
Oligonucleotide Microarray ◽  
Influenza Viruses ◽  
Isothermal Amplification ◽  
Detection Methods ◽  
Loop Mediated Isothermal Amplification ◽  
Viral Copy Number

H7N9 avian influenza viruses have circulated in the human population and poultry flocks in China since 2013. H7N9 virus monitoring is imperative in Taiwan due to the frequent contact between China and Taiwan. Traditional viral molecular detection methods using RT-PCR and sequencing are time- and labor-intensive, thus a simpler and cheaper method with high sensitivity and specificity is worth developing. We successfully detected human and wild bird H7N9 viruses in this study using reverse transcription loop-mediated isothermal amplification (RT-LAMP) and oligonucleotide microarray. The detection limit was as low as one viral copy number. The specific matching reaction between the templates and microarray probes during hybridization ensured the high detection effectiveness. The excellent sensitivity and specificity of the RT-LAMP-microarray makes it a powerful H7N9 surveillance approach in Taiwan.

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