scholarly journals Optimization of droplet digital PCR assays for the type specific detection and quantification of five HPV genotypes, also including additional data on viral load of nine different HPV genotypes in cervical carcinomas

2021 ◽  
pp. 114193
Author(s):  
Kaliff Malin ◽  
Bohr Mordhorst Louise ◽  
Helenius Gisela ◽  
Karlsson G. Mats ◽  
Lillsunde-Larsson Gabriella
Keyword(s):  
Viral Load ◽  
Additional Data ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Specific Detection ◽  
Cervical Carcinomas ◽  
Hpv Genotypes ◽  
Pcr Assays ◽  
Detection And Quantification

scholarly journals Detection and quantification of chimerism by droplet digital PCR

Chimerism ◽  
2013 ◽  
Vol 4 (3) ◽  
pp. 102-108 ◽  
Author(s):  
David George ◽  
Juliann Czech ◽  
Bobby John ◽  
Min Yu ◽  
Lawrence J. Jennings
Keyword(s):  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Detection And Quantification

scholarly journals Development and Evaluation of a Single Dye Duplex Droplet Digital PCR Assay for the Rapid Detection and Quantification of Mycobacterium tuberculosis

2020 ◽  
Vol 8 (5) ◽  
pp. 701 ◽  
Author(s):  
Raphael Nyaruaba ◽  
Jin Xiong ◽  
Caroline Mwaliko ◽  
Nuo Wang ◽  
Belindah J. Kibii ◽  
...  
Keyword(s):  
Target Genes ◽  
Annealing Temperature ◽  
Single Channel ◽  
Digital Pcr ◽  
Absolute Quantification ◽  
Droplet Digital Pcr ◽  
Sample Concentration ◽  
Specificity And Sensitivity ◽  
Probe Concentration ◽  
Detection And Quantification

Droplet digital PCR (ddPCR) is a third generation of PCR that was recently developed to overcome the challenges of real-time fluorescence-based quantitative PCR (qPCR) in absolute quantification of pathogens. Few studies have been done on tuberculosis (TB) detection and quantification using ddPCR despite its many advantages over qPCR. From the few studies, none explores a single dye duplex assay for the detection and quantification of TB. In this study, steps toward developing and evaluating a duplex single dye (FAM) assay for detecting two targets (IS6110 and IS1081) are clearly described using simplex and duplex experiments. To achieve this, various parameters are investigated, including annealing temperature, primer and probe concentration, sensitivity and specificity, sample concentration, and inter/intra-assay variability. From the results, primer and probe concentration, annealing temperature, and sample concentration have an effect on the position and separation of droplets in both simplex and duplex assays. The copies of target genes in a duplex assay can be estimated accurately using the threshold tool with little inter-assay (CV <1%) and intra-assay (CV <6%) variability when compared to simplex assays. The ddPCR assay specificity and sensitivity are both 100% when compared to qPCR. This work shows steps toward the detection and quantification of two targets in a single channel, enabling higher multiplexing to include more targets in future works.

scholarly journals Droplet digital PCR for detection and quantification of circulating tumor DNA in plasma of head and neck cancer patients

BMC Cancer ◽  
2017 ◽  
Vol 17 (1) ◽  
Author(s):  
Joost H. van Ginkel ◽  
Manon M. H. Huibers ◽  
Robert J. J. van Es ◽  
Remco de Bree ◽  
Stefan M. Willems
Keyword(s):  
Head And Neck Cancer ◽  
Head And Neck ◽  
Cancer Patients ◽  
Neck Cancer ◽  
Digital Pcr ◽  
Circulating Tumor Dna ◽  
Droplet Digital Pcr ◽  
Tumor Dna ◽  
Detection And Quantification

Sensitive and specific detection of lung cancer DNA in bronchial washing by a novel methylation-specific droplet digital PCR

2017 ◽  
Author(s):  
Gian Luca Casoni ◽  
Elena Miotto ◽  
Elena Saccenti ◽  
Susanna Mascetti ◽  
Debora Rasio ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Specific Detection ◽  
Bronchial Washing

scholarly journals Direct Quantitation of SARS-CoV-2 Using Droplet Digital PCR in Suspected Samples With Very Low Viral Load

2021 ◽  
Author(s):  
Michela Deiana ◽  
Chiara Piubelli ◽  
Antonio Mori ◽  
Gian Paolo Chiecchi ◽  
Giulia La Marca ◽  
...  
Keyword(s):  
Viral Load ◽  
Real Time ◽  
False Negative ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Hospitalized Patients ◽  
Lower Sensitivity ◽  
Rt Pcr ◽  
Viral Genes ◽  
Viral Target

Background: The reference test for SARS-CoV-2 detection is the reverse transcriptase real time PCR (real time RT-PCR). However, evidences reported that real time RT-PCR has a lower sensitivity compared with the droplet digital PCR (ddPCR) leading to possible false negative in low viral load cases. Methods: We used ddPCR for viral genes N1 and N2 on 20 negative (no detection) samples from symptomatic hospitalized COVID-patients presenting fluctuating real time RT-PCR results and 10 suspected samples (Ct value&gt;35) from asymptomatic not hospitalized subjects. Results: ddPCR performed on RNA revealed 65% of positivity for at least one viral target in the hospitalized patients group of samples (35% for N1 and N2, 10% only for N1 and 20% only for N2) and 50% in the suspected cases (30% for N1 and N2, while 20% only for N2). On hospitalized patients&rsquo; samples, we applied also a direct ddPCR approach on the swab material, achieving an overall positivity of 83%. Conclusion: ddPCR, in particular the direct quantitation on swabs, shows a sensitivity advantage for the SARS-CoV-2 identification and may be useful to reduce the false negative diagnosis, especially for low viral load suspected samples.

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