Droplet digital PCR for quantitative detection of Escherichia coli

2019 ◽  
Author(s):  
Jiayi Yang ◽  
Boqiang Fu ◽  
Chunyan Niu ◽  
Jing Wang
Keyword(s):  
Escherichia Coli ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Quantitative Detection

scholarly journals Development of a duplex droplet digital PCR assay for absolute quantitative detection of "Candidatus Liberibacter asiaticus"

PLoS ONE ◽  
2018 ◽  
Vol 13 (5) ◽  
pp. e0197184 ◽  
Author(s):  
Vijayanandraj Selvaraj ◽  
Yogita Maheshwari ◽  
Subhas Hajeri ◽  
Jianchi Chen ◽  
Thomas Greg McCollum ◽  
...  
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Quantitative Detection ◽  
Candidatus Liberibacter Asiaticus ◽  
Pcr Assay ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

scholarly journals Development and Validation of a Gamma Globin RT-Ddpcr Exploratory Biomarker for Sickle Cell Phase 1 Clinical Trial Fis 002-2020

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 4166-4166
Author(s):  
Mark Roth ◽  
Jian Yang ◽  
Dimitra Tsavachidou ◽  
Darren W. Davis
Keyword(s):  
Clinical Trial ◽  
Sickle Cell ◽  
Gamma Globulin ◽  
Phase 1 ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Quality Data ◽  
Quantitative Detection ◽  
Phase 1 Clinical Trial ◽  
Development And Validation

Abstract FTX-6058, a selective and potent binder of embryonic ectoderm development protein (EED), is being investigated in Sickle Cell Disease (SCD). In-vitro and pre-clinical Townes mouse studies show FTX-6058 upregulates expression of the gamma globulin gene leading to increased levels of fetal hemoglobin (HbF). In human clinical trials, as an early indicator of FTX-6058 induced gamma globulin expression, a reverse transcriptase droplet digital PCR (RT-ddPCR) assay was developed and validated by Precision for Medicine. Versus RT-qPCR, RT-ddPCR gives absolute copy number, does not require a standard curve, and is more precise and reproducible for low expressed targets 1,2. Precision for Medicine developed and validated RT-ddPCR assays for the target globin genes α, β, and γ and for the housekeeping genes TFRC and OAZ1. RNA isolated from 3 healthy and 3 SCD affected individuals was used for development and validation of the assay. The upper and lower limits of quantification were 120,000 and 10.5 copies/20μL reaction, respectively. For samples with >50 copies/20μL reaction, the CV for technical replicates was <20% for all transcripts. Furthermore, Precision for Medicine demonstrated SCD affected individuals express approximately 15-fold more γ globulin versus non affected individuals. Fulcrum utilized the globin RT-ddPCR assay validated by Precision for Medicine in the FTX-6058 phase 1 clinical trial FIS 002-2020. 1 Zhao Y, Xia Q, Yin Y, Wang Z (2016), Comparison of Droplet Digital PCR and Quantitative PCR Assays for Quantitative Detection of Xanthomonas citri Subsp. citri. PLoS ONE 11(7): e0159004. doi:10.1371/journal.pone.0159004 2 Taylor SC, Laperriere G, Germain H (2017), Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data. Scientific Reports 7(2409): DOI:10.1038/s41598-017-02217-x Disclosures Roth: Fulcrum Therapeutics, Inc.: Current Employment, Current equity holder in publicly-traded company. Yang: Fulcrum Therapeutics, Inc.: Consultancy. Tsavachidou: Fulcrum Therapeutics, Inc.: Consultancy. Davis: Fulcrum Therapeutics, Inc.: Consultancy.

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