Digital Droplet PCR Method for the Quantification of AAV Transduction Efficiency in Murine Retina

10.3791/63038 ◽  
2021 ◽  
Author(s):  
Iskalen Cansu Topcu Okan ◽  
Mehri Ahmadian ◽  
Yesim Tutuncu ◽  
Halit Yusuf Altay ◽  
Cavit Agca
Keyword(s):  
Transduction Efficiency ◽  
Digital Droplet Pcr ◽  
Droplet Pcr ◽  
Pcr Method

scholarly journals Monitoring EGFR C797S mutation in Japanese NSCLC patients with serial cell free DNA evaluation using digital droplet PCR

2021 ◽  
Author(s):  
Ryo Ariyasu ◽  
Ken Uchibori ◽  
Takaaki Sasaki ◽  
Mika Tsukahara ◽  
Kazuma Kiyotani ◽  
...  
Keyword(s):  
Cell Free Dna ◽  
Digital Droplet Pcr ◽  
Free Dna ◽  
Droplet Pcr ◽  
Nsclc Patients

scholarly journals P34.06 The Clinical Utility of Liquid Biopsy by Digital Droplet PCR in Patients with Advanced NSCLC

2021 ◽  
Vol 16 (3) ◽  
pp. S417
Author(s):  
V. Lamberts ◽  
M. Aldea ◽  
L. Mezquita ◽  
C. Jovelet ◽  
D. Vasseur ◽  
...  
Keyword(s):  
Liquid Biopsy ◽  
Clinical Utility ◽  
Advanced Nsclc ◽  
Digital Droplet Pcr ◽  
Droplet Pcr

Carbon metabolism snapshot by ddPCR during the early step of Candida albicans phagocytosis by macrophages

2020 ◽  
Vol 78 (1) ◽  
Author(s):  
Romain Laurian ◽  
Cécile Jacot-des-Combes ◽  
Fabiola Bastian ◽  
Karine Dementhon ◽  
Pascale Cotton
Keyword(s):  
Carbon Metabolism ◽  
Early Stage ◽  
Yeast Cells ◽  
Transcription Level ◽  
Digital Droplet Pcr ◽  
Metabolic Genes ◽  
Glucose Transport System ◽  
Short Period ◽  
Droplet Pcr ◽  
Alternative Sources

ABSTRACT During Candida macrophage interactions, phagocytosed yeast cells feed in order to grow, develop hyphae and escape. Through numerous proteomic and transcriptomic studies, two metabolic phases have been described. A shift to a starvation mode is generally identified as early as one-hour post phagocytosis, followed by a glycolytic growth mode after C. albicans escaped from the macrophage. Healthy macrophages contain low amounts of glucose. To determine if this carbon source was sensed and metabolized by the pathogen, we explored the transcription level of a delimited set of key genes expressed in C. albicans cells during phagocytosis by macrophages, at an early stage of the interaction. This analysis was performed using a technical digital droplet PCR approach to quantify reliably the expression of carbon metabolic genes after 30 min of phagocytosis. Our data confirm the technique of digital droplet PCR for the detection of C. albicans transcripts using cells recovered after a short period of phagocytosis. At this stage, carbon metabolism is clearly oriented towards the use of alternative sources. However, the activation of high-affinity glucose transport system suggests that the low amount of glucose initially present in the macrophages is detected by the pathogen.

scholarly journals Quantification of SARS-CoV-2 viral copy number in saliva mouthwash samples using digital droplet PCR

2020 ◽  
Author(s):  
Lisa Oberding ◽  
Jia Hu ◽  
Byron Berenger ◽  
Abu Naser Mohon ◽  
Dylan R. Pillai
Keyword(s):  
Viral Load ◽  
Copy Number ◽  
Clinical Samples ◽  
Digital Droplet Pcr ◽  
Viral Copy Number ◽  
Droplet Pcr ◽  
Precise Quantification ◽  
Viral Copy ◽  
Mouth Wash

AbstractSaliva samples were collected through a simple mouth wash procedure and viral load quantified using a technology called digital droplet PCR. Data suggest ddPCR allows for precise quantification of viral load in clinical samples infected with SARS-CoV-2.

scholarly journals Digital Droplet PCR: CNV Analysis and Other Applications

2014 ◽  
Vol 82 (1) ◽  
Author(s):  
Erica Mazaika ◽  
Jason Homsy
Keyword(s):  
Digital Droplet Pcr ◽  
Droplet Pcr

Digital Droplet PCR (ddPCR) for Detection and Quantitation of Hepatitis Delt Virus

2021 ◽  
Author(s):  
Ling Xu ◽  
Xiangying Zhang ◽  
Yaling Cao ◽  
Zihao Fan ◽  
Yuan Tian ◽  
...  
Keyword(s):  
Liver Failure ◽  
Lower Limit ◽  
Limit Of Detection ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Rna Detection ◽  
Detection Rates ◽  
Digital Droplet Pcr ◽  
Droplet Pcr ◽  
Hdv Infection

Abstract Background & Aims The prevalence of hepatitis delt virus (HDV) far exceeds our expected level, there remains a lack of reliable quantitative assays for HDV RNA detection. We sought to develop a new method based on digital droplet PCR (ddPCR) for HDV RNA quantitative detection. Methods With plasmid (pMD19T) containing HDV full-genome, we determined the method for ddPCR-based HDV RNA quantification. To compare various assays for HDV detection, 30 cases diagnosed hepatitis D and 14 controls were examined by ELISA, RT-PCR and ddPCR. 728 HBV-related patients including 182 chronic hepatitis B (CHB), 182 liver cirrhosis (LC), 182 hepatocellular carcinoma (HCC) and 182 liver failure (LF) were screened for HDV infection. Results The limit of detection of ddPCR for HDV is significantly low, which lower limit of detection (LLoD) and lower limit of quantitation (LLoQ) to be 5.51 copies/reaction (95% CI: 1.15–6.4*105) and 0.18 copies/reaction (95% CI: 0.0012151- 0.76436), respectively. Among the 44 samples, ELISA detected 30 cases positive for anti-HDV, ddPCR reported 24 samples and RT-PCR reported 10 samples positive for HDV RNA. Moreover, the positive rates of anti-HDV IgG were 1.1%, 3.3%, 2.7% and 7.1% in patients with CHB, LC, HCC, and LF; the detection rates of RT-PCR in HDV RNA were 0%, 16.67%, 15.4% and 20%, however, the detection rates of ddPCR were 0%, 33.33%, 30.77% and 60%. Conclusion We establish a high sensitivity and high specificity quantitative HDV RNA detection method based on ddPCR compared to RT-PCR. HBV-related end-stage liver disease, especially liver failure, are associated with a remarkably high rate of HDV infection.

Development of quantitative real-time PCR and digital droplet-PCR assays for rapid and early detection of the spoilage yeasts Saccharomycopsis fibuligera and Wickerhamomyces anomalus in bread

2022 ◽  
Vol 101 ◽  
pp. 103894
Author(s):  
Paola Cremonesi ◽  
Cristiana Garofalo ◽  
Claudia Picozzi ◽  
Bianca Castiglioni ◽  
Nicola Mangieri ◽  
...  
Keyword(s):  
Early Detection ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Saccharomycopsis Fibuligera ◽  
Wickerhamomyces Anomalus ◽  
Digital Droplet Pcr ◽  
Spoilage Yeasts ◽  
Droplet Pcr ◽  
Pcr Assays

Quantification of the allergen soy (Glycine max) in food using digital droplet PCR (ddPCR)

2018 ◽  
Vol 245 (2) ◽  
pp. 499-509 ◽  
Author(s):  
W. Mayer ◽  
M. Schuller ◽  
M. C. Viehauser ◽  
R. Hochegger
Keyword(s):  
Glycine Max ◽  
Digital Droplet Pcr ◽  
Droplet Pcr
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