scholarly journals A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach

2013 ◽  
pp. n/a-n/a
Author(s):  
Y. Yu ◽  
Z. Zhao ◽  
D. Jiang ◽  
Z. Wu ◽  
S. Li
Keyword(s):  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

A one-step multiplex real-time RT-PCR assay for rapid and simultaneous detection of human norovirus genogroup I, II and IV

2013 ◽  
Vol 189 (2) ◽  
pp. 277-282 ◽  
Author(s):  
Yong Yan ◽  
Heng-hui Wang ◽  
Lei Gao ◽  
Ji-mei Ji ◽  
Zhi-jie Ge ◽  
...  
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Human Norovirus ◽  
One Step

scholarly journals Development and evaluation of one-step multiplex real-time RT-PCR assay for simultaneous detection of Zika virus and Chikungunya virus

2017 ◽  
Vol 90 (3) ◽  
pp. 389-396 ◽  
Author(s):  
Si-Qing Liu ◽  
Xiao Li ◽  
Cheng-Lin Deng ◽  
Zhi-Ming Yuan ◽  
Bo Zhang
Keyword(s):  
Real Time ◽  
Zika Virus ◽  
Chikungunya Virus ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

scholarly journals Development of a multiplex one-step real-time RT-PCR assay for the simultaneous detection of eight viruses associated with febrile rash illnesses

2020 ◽  
Vol 2 (2) ◽  
pp. 89-94 ◽  
Author(s):  
Aili Cui ◽  
Shulei Wang ◽  
Qiang Zhang ◽  
Huiling Wang ◽  
Zhen Zhu ◽  
...  
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

scholarly journals Sensitive Detection and Quantification of SARS-CoV-2 by Multiplex Droplet Digital RT-PCR

2020 ◽  
Author(s):  
Remco de Kock ◽  
Mieke Baselmans ◽  
Volkher Scharnhorst ◽  
Birgit Deiman
Keyword(s):  
Simultaneous Detection ◽  
Housekeeping Gene ◽  
Multiplex Assay ◽  
Viral Rna ◽  
Sensitive Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Assay Sensitivity ◽  
One Step ◽  
Detection And Quantification

Abstract Purpose. We aimed to develop a one-step droplet digital RT-PCR (RT-ddPCR) multiplex assay that allows for sensitive detection of SARS-CoV-2 RNA with respect to human derived RNA and could be used for triage and monitoring of Covid-19 patients. Methods. A one step RT-ddPCR multiplex assay was developed for simultaneous detection of SARS-CoV-2 E, RdRp and N viral RNA, and human Rpp30 DNA and GUSB mRNA, for internal nucleic acid (NA) extraction and RT-PCR control. Dilution series of viral RNA transcripts were prepared in water and total NA extract of Covid-19 negative patients. As reference assay, an E-GUSB duplex RT-PCR was used. Results. Assay sensitivity of the RT-PCR assay drastically decreased when SARS-CoV-2 copies were detected in a background of total NA extract compared to water, while the sensitivity of the RT-ddPCR was not affected by the total NA background. GUSB mRNA detection was used to set validity criteria to assure viral RNA and RT-PCR assay quality. In a background of at least 100 GUSB mRNA copies, 5 copies of viral RNA are reliably detectable and 10 copies viral RNA copies are reliably quantifiable. Conclusion. The present study describes a robust and sensitive one-step RT-ddPCR multiplex assay for reliable detection and quantification of SARS-CoV-2 RNA. By determining the fractional abundance of viral RNA with respect to a human housekeeping gene, viral loads from different samples can be compared, what could be used to investigate the infectiveness and to monitor Covid-19 patients.

scholarly journals Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR

2020 ◽  
Author(s):  
Remco de Kock ◽  
Mieke Baselmans ◽  
Volkher Scharnhorst ◽  
Birgit Deiman
Keyword(s):  
Simultaneous Detection ◽  
Housekeeping Gene ◽  
Multiplex Assay ◽  
Viral Rna ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Assay Sensitivity ◽  
Viral Loads ◽  
Pcr Multiplex ◽  
One Step

Abstract The purpose of this study is to develop a one-step droplet digital RT-PCR (RT-ddPCR) multiplex assay that allows for sensitive quantification of SARS-CoV-2 RNA with respect to human-derived RNA and could be used for screening and monitoring of Covid-19 patients. A one-step RT-ddPCR multiplex assay was developed for simultaneous detection of SARS-CoV-2 E, RdRp and N viral RNA, and human Rpp30 DNA and GUSB mRNA, for internal nucleic acid (NA) extraction and RT-PCR control. Dilution series of viral RNA transcripts were prepared in water and total NA extract of Covid-19-negative patients. As reference assay, an E-GUSB duplex RT-PCR was used. GUSB mRNA detection was used to set validity criteria to assure viral RNA and RT-PCR assay quality and to enable quantification of SARS-CoV-2 RNA. In a background of at least 100 GUSB mRNA copies, 5 copies of viral RNA are reliably detectable and 10 copies viral RNA copies are reliably quantifiable. It was found that assay sensitivity of the RT-ddPCR was not affected by the total NA background while assay sensitivity of the gold standard RT-PCR assay is drastically decreased when SARS-CoV-2 copies were detected in a background of total NA extract compared with water. The present study describes a robust and sensitive one-step ddRT-PCR multiplex assay for reliable quantification of SARS-CoV-2 RNA. By determining the fractional abundance of viral RNA with respect to a human housekeeping gene, viral loads from different samples can be compared, what could be used to investigate the infectiveness and to monitor Covid-19 patients.

A rapid one-step multiplex RT-PCR assay for the simultaneous detection of five citrus viroids in China

2008 ◽  
Vol 124 (1) ◽  
pp. 175-180 ◽  
Author(s):  
Xuefeng Wang ◽  
Changyong Zhou ◽  
Kezhi Tang ◽  
Yan Zhou ◽  
Zhongan Li
Keyword(s):  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Citrus Viroids ◽  
One Step

scholarly journals Surveillance of dengue virus in individual Aedes aegypti mosquitoes collected concurrently with suspected human cases in Tarlac City, Philippines

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Jean Claude Balingit ◽  
Thaddeus M. Carvajal ◽  
Mariko Saito-Obata ◽  
Maribet Gamboa ◽  
Amalea Dulcene Nicolasora ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Vector Control ◽  
Real Time ◽  
Phylogenetic Analyses ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Denv Serotypes ◽  
One Step

Abstract Background Vector control measures are critical for the prevention and reduction of dengue virus (DENV) transmission. Effective vector control is reliant not only on knowledge of mosquito abundance, but also on the timely and accurate detection of mosquito-borne infection. Mosquito-based virus surveillance programs typically rely on pool-based mosquito testing, although whether individual-based mosquito testing is a feasible alternative to this has not been widely studied. Applying an individual-based mosquito testing approach, we conducted a 1-month surveillance study of DENV in adult Aedes aegypti mosquitoes in homes of suspected dengue patients during the 2015 peak dengue season in Tarlac City, Philippines to more accurately assess the mosquito infection rate and identify the DENV serotypes and genotypes concurrently co-circulating in mosquitoes and patients there. Methods We performed a one-step multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR) assay for the simultaneous detection and serotyping of DENV in patients and individual female Ae. aegypti mosquitoes. Additionally, we performed sequencing and phylogenetic analyses to further characterize the detected DENV serotypes in mosquitoes and patients at the genotype level. Results We collected a total of 583 adult Ae. aegypti mosquitoes, of which we individually tested 359 female mosquitoes for the presence of DENV. Ten (2.8%) of the 359 female mosquitoes were positive for the presence of DENV. We detected DENV-1, DENV-2, and DENV-4 in the field-collected mosquitoes, which was consistent with the serotypes concurrently found in infected patients. Sequencing and phylogenetic analyses of the detected DENV serotypes based on the partial sequence of the evelope (E) gene revealed three genotypes concurrently present in the sampled mosquitoes and patients during the study period, namely DENV-1 genotype IV, DENV-2 Cosmopolitan genotype, and DENV-4 genotype II. Conclusions We demonstrated the utility of a one-step multiplex real-time RT-PCR assay for the individual-based DENV surveillance of mosquitoes. Our findings reinforce the importance of detecting and monitoring virus activity in local mosquito populations, which are critical for dengue prevention and control.

scholarly journals A One-Step, Triplex, Real-Time RT-PCR Assay for the Simultaneous Detection of Enterovirus 71, Coxsackie A16 and Pan-Enterovirus in a Single Tube

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e102724 ◽  
Author(s):  
Shiyin Zhang ◽  
Jin Wang ◽  
Qiang Yan ◽  
Shuizhen He ◽  
Wenbin Zhou ◽  
...  
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Enterovirus 71 ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Single Tube ◽  
One Step
Sign in / Sign up

Export Citation Format

Share Document