Development and application of reverse transcription droplet digital PCR assay for sensitive detection of apple scar skin viroid during in vitro propagation of apple plantlets

2021 ◽  
pp. 101789
Author(s):  
Hyo-Jeong Lee ◽  
Yeon Soo Han ◽  
In-Sook Cho ◽  
Rae-Dong Jeong
Keyword(s):  
Reverse Transcription ◽  
In Vitro Propagation ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Pcr Assay ◽  
Scar Skin

Development of a novel reverse transcription droplet digital PCR assay for the sensitive detection of Senecavirus A

2018 ◽  
Vol 66 (1) ◽  
pp. 517-525 ◽  
Author(s):  
Zhou Zhang ◽  
Yongning Zhang ◽  
Xiangmei Lin ◽  
Zhenhai Chen ◽  
Shaoqiang Wu
Keyword(s):  
Reverse Transcription ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Pcr Assay

Development of a sensitive and reliable reverse transcription droplet digital PCR assay for the detection of citrus yellow vein clearing virus

2018 ◽  
Vol 164 (3) ◽  
pp. 691-697 ◽  
Author(s):  
Yingjie Liu ◽  
Yingli Wang ◽  
Qin Wang ◽  
Yanhui Zhang ◽  
Wanxia Shen ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Yellow Vein ◽  
Pcr Assay ◽  
Vein Clearing

scholarly journals Development of a Multiplex Droplet Digital PCR Assay for the Detection of Babesia, Bartonella, and Borrelia Species

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1462
Author(s):  
Ricardo Maggi ◽  
Edward B. Breitschwerdt ◽  
Barbara Qurollo ◽  
Jennifer C. Miller
Keyword(s):  
Animal Cell ◽  
Simultaneous Detection ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Pcr Assay ◽  
Theileria Equi ◽  
Cytauxzoon Felis ◽  
Vector Borne ◽  
Type Water

We describe the development, optimization, and validation of a multiplex droplet digital PCR (ddPCR) assay for the simultaneous detection of Babesia, Bartonella, and Borrelia spp. DNA from several sample matrices, including clinical blood samples from animals and humans, vectors, in-vitro infected human and animal cell lines, and tissues obtained from animal models (infected with Bartonella and/or B. burgdorferi). The multiplex ddPCR assay was able to detect 31 Bartonella, 13 Borrelia, and 24 Babesia species, including Theileria equi, T. cervi, and Cytauxzoon felis. No amplification of Treponema or Leptospira spp. was observed. Sensitivity of 0.2–5 genome equivalent DNA copies per microliter was achieved for different members of the Bartonella and Borrelia genus, depending on the species or matrix type (water or spiked blood DNA) tested. The ddPCR assay facilitated the simultaneous detection of co-infections with two and three vector-borne pathogens comprising four different genera (Babesia, Bartonella, Borrelia, and Theileria) from clinical and other sample sources.

Abstract 411: Rapid and sensitive detection of EGFR C797S mutations using a blood-based droplet digital PCR assay

2019 ◽  
Author(s):  
Victoria Edwards ◽  
Leisa Jackson ◽  
Jordan Reese ◽  
Hestia S. Mellert ◽  
Gary Pestano
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Pcr Assay

scholarly journals Developing multiplex ddPCR assays for SARS-CoV-2 detection based on probe mix and amplitude based multiplexing

2020 ◽  
Author(s):  
Raphael Nyaruaba ◽  
Changchang Li ◽  
Caroline Mwaliko ◽  
Matilu Mwau ◽  
Nelson Odiwour ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Digital Pcr ◽  
Multiplex Assay ◽  
Small Sample ◽  
Droplet Digital Pcr ◽  
Rt Pcr ◽  
Multiplex Assays ◽  
Reverse Transcription Pcr ◽  
Sample Throughput

AbstractMultiplexing has been highlighted to save on costs, increase sample throughput, and maximize on the number of targets that can be sensitively detected within a small sample. With the ongoing SARS-CoV-2 pandemic, different articles have been published highlighting the superiority of droplet digital PCR (ddPCR) over the gold reverse transcription PCR (RT-PCR) in SARS-CoV-2 detection. However, few studies have been reported on developing multiplex ddPCR assays for SARS-CoV-2 detection and their performance. In this study, we developed simplex (1 target), duplex (2 targets), triplex probe mix (3 targets), and fourplex (4 targets) assays based on a two color ddPCR system for SARS-CoV-2 detection. Results showed that the fourplex assay had the similar limits of detection and accuracy to the lower multiplex assays. Analyzing 94 clinical isolates demonstrated that the ddPCR triplex probe mix assay had better sensitivity than the RT-qPCR assay. Additionally, the ddPCR multiplex assay showed that remdesivir could inhibit the growth of SARS-CoV-2 in vitro while another testing drug couldn’t. Conclusively, our research shows that developing multiplex ddPCR assays is possible by combing probe mix and amplitude based multiplexing, which will help in developing multiplexed ddPCR assays for different SARS-CoV-2 applications.

Development of a droplet digital PCR assay for sensitive detection of porcine circovirus 3

2019 ◽  
Vol 43 ◽  
pp. 50-57 ◽  
Author(s):  
Yongning Zhang ◽  
Zhou Zhang ◽  
Zhanying Wang ◽  
Zili Wang ◽  
Caixia Wang ◽  
...  
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Porcine Circovirus ◽  
Pcr Assay ◽  
Porcine Circovirus 3

Development of a novel droplet digital PCR assay for the sensitive detection of carp edema virus

Aquaculture ◽  
2021 ◽  
pp. 737162
Author(s):  
Na Wang ◽  
Min Zhang ◽  
Zhou Zhang ◽  
Hongli Jing ◽  
Shaoqiang Wu ◽  
...  
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Pcr Assay

Abstract 411: Rapid and sensitive detection of EGFR C797S mutations using a blood-based droplet digital PCR assay

2019 ◽  
Author(s):  
Victoria Edwards ◽  
Leisa Jackson ◽  
Jordan Reese ◽  
Hestia S. Mellert ◽  
Gary Pestano
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Pcr Assay

scholarly journals Development of a reverse transcription droplet digital PCR (RT-ddPCR) assay for sensitive detection of simian immunodeficiency virus (SIV)

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Samuel Long ◽  
Brian Berkemeier
Keyword(s):  
Simian Immunodeficiency Virus ◽  
Reverse Transcription ◽  
Rhesus Macaques ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Sensitive Detection ◽  
Similar Reaction ◽  
Tissue Samples ◽  
Low Level ◽  
Immunodeficiency Virus

Abstract Background Simian immunodeficiency virus (SIV)-infected rhesus macaques constitute an excellent model of human HIV infection. Sensitive detection of SIV RNA in cell and tissue samples from infected animals subjected to treatment regimens becomes especially critical in determining which therapeutic attempts are successful, and consequently, which interventions should be prioritized in HIV cure research. Results In this report, we describe the design and testing of a Raindance ddPCR platform-based, sensitive SIV reverse transcription droplet digital PCR (RT-ddPCR) assay by exploring the combinations of various priming conditions and reverse transcriptases, and testing one-step vs. two-step procedures, to eliminate background signal(s) and enable detection and quantification of low level target signals. Conclusions Similar reaction conditions and assay validation procedures can be explored for potential development of additional assays for other applications that require sensitive detection of low-level targets in RNA samples.

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