scholarly journals Application of droplet digital PCR to detect the pathogens of infectious diseases

2018 ◽  
Vol 38 (6) ◽  
Author(s):  
Haiyi Li ◽  
Ruolan Bai ◽  
Zhenyu Zhao ◽  
Lvyan Tao ◽  
Mingbiao Ma ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Infectious Diseases ◽  
Quantitative Polymerase Chain Reaction ◽  
Digital Pcr ◽  
Conventional Polymerase Chain Reaction ◽  
Nucleic Acid Detection ◽  
Chain Reaction ◽  
Molecular Biology Technique ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Polymerase chain reaction (PCR) is a molecular biology technique used to multiply certain deoxyribonucleic acid (DNA) fragments. It is a common and indispensable technique that has been applied in many areas, especially in clinical laboratories. The third generation of polymerase chain reaction, droplet digital polymerase chain reaction (ddPCR), is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify DNA. Droplet digital polymerase chain reaction is now widely used in low-abundance nucleic acid detection and is useful in diagnosis of infectious diseases. Here, we summarized the potential advantages of droplet digital polymerase chain reaction in clinical diagnosis of infectious diseases, including viral diseases, bacterial diseases and parasite infections, concluded that ddPCR provides a more sensitive, accurate, and reproducible detection of low-abundance pathogens and may be a better choice than quantitative polymerase chain reaction for clinical applications in the future.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Digital PCR: A Reliable Tool for Analyzing and Monitoring Hematologic Malignancies

2020 ◽  
Vol 21 (9) ◽  
pp. 3141 ◽  
Author(s):  
Nicoletta Coccaro ◽  
Giuseppina Tota ◽  
Luisa Anelli ◽  
Antonella Zagaria ◽  
Giorgina Specchia ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Residual Disease ◽  
Hematologic Malignancies ◽  
Digital Pcr ◽  
Future Perspectives ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction ◽  
Generation Sequencing ◽  
Minimal Residual

The digital polymerase chain reaction (dPCR) is considered to be the third-generation polymerase chain reaction (PCR), as it yields direct, absolute and precise measures of target sequences. dPCR has proven particularly useful for the accurate detection and quantification of low-abundance nucleic acids, highlighting its advantages in cancer diagnosis and in predicting recurrence and monitoring minimal residual disease, mostly coupled with next generation sequencing. In the last few years, a series of studies have employed dPCR for the analysis of hematologic malignancies. In this review, we will summarize these findings, attempting to focus on the potential future perspectives of the application of this promising technology.

Heterogeneous modification of through-hole microwell chips for ultralow cross-contamination digital polymerase chain reaction

The Analyst ◽  
2020 ◽  
Vol 145 (8) ◽  
pp. 3116-3124
Author(s):  
Jinze Li ◽  
Yajun Qiu ◽  
Zhiqi Zhang ◽  
Chuanyu Li ◽  
Shuli Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Digital Pcr ◽  
Cross Contamination ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction ◽  
Through Hole

Heterogeneous modification of through-hole microwell chips to avoid cross-contamination during digital PCR.

A localized temporary negative pressure assisted microfluidic device for detecting keratin 19 in A549 lung carcinoma cells with digital PCR

2015 ◽  
Vol 7 (5) ◽  
pp. 2006-2011 ◽  
Author(s):  
Qingchang Tian ◽  
Qi Song ◽  
Yanan Xu ◽  
Qiangyuan Zhu ◽  
Bingwen Yu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Digital Pcr ◽  
Carcinoma Cells ◽  
Biological Research ◽  
Chain Reaction ◽  
Lung Carcinoma Cells ◽  
Keratin 19 ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction ◽  
A549 Lung

Digital polymerase chain reaction (dPCR) has played a major role in biological research, especially by providing an accurate counting of single nucleic acid molecules.

Massive droplets generation for digital PCR via a smart step emulsification chip integrated in a reaction tube

The Analyst ◽  
2021 ◽  
Author(s):  
Shuhao Zhao ◽  
Zengming Zhang ◽  
Fei Hu ◽  
Junjun Wu ◽  
Niancai Peng
Keyword(s):  
Polymerase Chain Reaction ◽  
Large Scale ◽  
Digital Pcr ◽  
Chain Reaction ◽  
Reaction Tube ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction ◽  
Monodisperse Droplets

Step emulsification (SE) devices coupled with parallel generation nozzles is widely used in the production of large-scale monodisperse droplets, especially for droplet based digital polymerase chain reaction (ddPCR) analysis. Although...

scholarly journals Digital polymerase chain reaction in an array of microfluidic printed droplets

2019 ◽  
Author(s):  
Yongfan Men ◽  
Jiannan Li ◽  
Tingting Ao ◽  
Zhihao Li ◽  
Bizhu Wu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Cost Effective ◽  
Digital Pcr ◽  
Research Field ◽  
Clinical Diagnostics ◽  
Future Market ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
New Perspective ◽  
Digital Polymerase Chain Reaction

AbstractDigital polymerase chain reaction (PCR) is a fast-developed technology, which makes it possible to provide absolute quantitative results. However, this technology has not been widely used in research field or clinical diagnostics. Although digital PCR has been born for two decades, the products on this subject still suffer from either high cost or cumbersome user experience, hence very few labs have the willingness or budget to routinely use such product; On the other hand, the unique sensitivity of dPCR over traditional qPCR shows great potential applications. Here, a cost-effective digital PCR method based on a microfluidic printing system was introduced, trying to overcome those shortcomings. The microfluidic droplet printing technology was utilized in this study to directly generate droplet array containing PCR reaction solution onto the simple glass substrate for the subsequent PCR and imaging, which could be done with any regular flat-panel PCR machine and microscope. The method introduces a new perspective in droplet-based digital PCR in that the droplets generated with this method aligns well in an array without touch with each other, therefore the regular glass and oil could be used without any special surfactant. With simple analysis, the data generated with this method showed reliable quality, which followed the Poisson distribution trend. Compared with other expensive digital PCR methods, this system is more affordable and simpler to integrate, especially for those biological or medical labs which are in need for the digital PCR options but short in budget. Therefore, this method is believed to have the great potential in the future market application.

scholarly journals Competitiveness of Quantitative Polymerase Chain Reaction (qPCR) and Droplet Digital Polymerase Chain Reaction (ddPCR) Technologies, with a Particular Focus on Detection of Antibiotic Resistance Genes (ARGs)

2021 ◽  
Vol 1 (3) ◽  
pp. 426-444
Author(s):  
Sol Park ◽  
Anita Rana ◽  
Way Sung ◽  
Mariya Munir
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
Viral Infectious Disease ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

With fast-growing polymerase chain reaction (PCR) technologies and various application methods, the technique has benefited science and medical fields. While having strengths and limitations on each technology, there are not many studies comparing the efficiency and specificity of PCR technologies. The objective of this review is to summarize a large amount of scattered information on PCR technologies focused on the two majorly used technologies: qPCR (quantitative polymerase chain reaction) and ddPCR (droplet-digital polymerase chain reaction). Here we analyze and compare the two methods for (1) efficiency, (2) range of detection and limitations under different disciplines and gene targets, (3) optimization, and (4) status on antibiotic resistance genes (ARGs) analysis. It has been identified that the range of detection and quantification limit varies depending on the PCR method and the type of sample. Careful optimization of target gene analysis is essential for building robust analysis for both qPCR and ddPCR. In our era where mutation of genes may lead to a pandemic of viral infectious disease or antibiotic resistance-induced health threats, this study hopes to set guidelines for meticulous detection, quantification, and analysis to help future prevention and protection of global health, the economy, and ecosystems.

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