Droplet Digital PCR Measurement of HER2 Copy Number Alteration in Formalin-Fixed Paraffin-Embedded Breast Carcinoma Tissue

Belgrader, Phillip; Tanner, Steven F.; Regan, John F.; Koehler, Ryan T.; Hindson, Benjamin J.; Brown, Alexandra S.

doi:10.1373/clinchem.2012.197855

Cited by 69 publications

(51 citation statements)

References 18 publications

(13 reference statements)

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“…It has the potential to have a major impact on molecular analyses ranging from clinical applications, such as biomarker analysis (2 ), viral detection (3 ), prognostic monitoring (4 ), and fetal screening (5 ), to research applications such as phage-host interactions (6 ) and intracellular profiling (7 ). dPCR can also be applied to assist with the library preparation needed for massively parallel (or next generation) sequencing methods (8 ).…”

confidence: 99%

The Digital MIQE Guidelines: Minimum Information for Publication of Quantitative Digital PCR Experiments

Huggett¹,

Foy²,

Beneš³

et al. 2013

Clinical Chemistry

738

663

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There is growing interest in digital PCR (dPCR) because technological progress makes it a practical and increasingly affordable technology. dPCR allows the precise quantification of nucleic acids, facilitating the measurement of small percentage differences and quantification of rare variants. dPCR may also be more reproducible and less susceptible to inhibition than quantitative real-time PCR (qPCR). Consequently, dPCR has the potential to have a substantial impact on research as well as diagnostic applications. However, as with qPCR, the ability to perform robust meaningful experiments requires careful design and adequate controls. To assist independent evaluation of experimental data, comprehensive disclosure of all relevant experimental details is required. To facilitate this process we present the Minimum Information for Publication of Quantitative Digital PCR Experiments guidelines. This report addresses known requirements for dPCR that have already been identified during this early stage of its development and commercial implementation. Adoption of these guidelines by the scientific community will help to standardize experimental protocols, maximize efficient utilization of resources, and enhance the impact of this promising new technology.

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confidence: 99%

The Digital MIQE Guidelines: Minimum Information for Publication of Quantitative Digital PCR Experiments

Huggett¹,

Foy²,

Beneš³

et al. 2013

Clinical Chemistry

738

663

View full text Add to dashboard Cite

show abstract

“…However, falsenegative results are common with these methods because of HBV surface antigen (HBsAg) variation and low HBV copy number. In particular, the precision limitations of qPCR have prevented distinguishing between small differences in copy number among samples (12 ), especially in cases of low copy number.…”

confidence: 99%

Next Generation Digital PCR Measurement of Hepatitis B Virus Copy Number in Formalin-Fixed Paraffin-Embedded Hepatocellular Carcinoma Tissue

et al. 2015

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BACKGROUND: Hepatocellular carcinoma (HCC) is strongly associated with hepatitis B virus (HBV) infection. False-negative results are common in routine serological tests and quantitative real-time PCR because of HBV surface antigen (HBsAg) variation and low HBV copy number. Droplet digital PCR (ddPCR), a next generation digital PCR, is a novel, sensitive, and specific platform that can be used to improve HBV detection.

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“…Furthermore, the variation in the expression of the HBV surface antigen (HBsAg) and low copy number of HBV underlie the frequent false negative results obtained by qPCR. Worst of all, the lack of precision in qPCR assays has prevented the detection of very small differences in copy number between samples, 7) especially when the initial copy numbers are already low. It is difficult to evaluate the results of qPCR when the Cq value of an unknown sample is close to the cutoff.…”

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confidence: 99%

Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA

Tang

Cai

et al. 2016

Bioscience, Biotechnology, and Biochemistry

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Quantitative real-time PCR (qPCR) has been widely implemented for clinical hepatitis B viral load testing, but a lack of standardization and relatively poor precision hinder its usefulness. Droplet digital PCR (ddPCR) is a promising tool that offers high precision and direct quantification. In this study, we compared the ddPCR QX100 platform by Bio-Rad with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad, USA) to detect serial plasmid DNA dilutions of known concentrations as well as HBV DNA extracted from patient serum samples. Both methods showed a high degree of linearity and quantitative correlation. However, ddPCR assays generated more reproducible results and detected lower copy numbers than qPCR assays. Patient sample quantifications by ddPCR and qPCR were highly agreeable based on the Bland-Altman analysis. Collectively, our findings demonstrate that ddPCR offers improved analytical sensitivity and specificity for HBV measurements and is suitable for clinical HBV detection.

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Droplet Digital PCR Measurement of HER2 Copy Number Alteration in Formalin-Fixed Paraffin-Embedded Breast Carcinoma Tissue

Cited by 69 publications

References 18 publications

The Digital MIQE Guidelines: Minimum Information for Publication of Quantitative Digital PCR Experiments

The Digital MIQE Guidelines: Minimum Information for Publication of Quantitative Digital PCR Experiments

Next Generation Digital PCR Measurement of Hepatitis B Virus Copy Number in Formalin-Fixed Paraffin-Embedded Hepatocellular Carcinoma Tissue

Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA

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