Species identification and quantification of silver pomfret using the droplet digital PCR assay

Cao, Weiwei; Li, Yiming; Chen, Xun; Chang, Yanlei; Li, Lili; Shi, Lei; Bai, Weibin; Ye, Lei

doi:10.1016/j.foodchem.2019.125331

Cited by 24 publications

(13 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…9 Additionally, the ddPCR system quantified the DNA in a highly reproducible manner without 11.0 ND 4 × 10 7 14.7 ND 4 × 10 6 18.2 100,000 4 relying on a standard curve. 2,8 However, based on analyzing 96 samples, the ddPCR took 3.3 times longer to complete than the rtPCR, the operation of ddPCR was more complicated than the rtPCR, and the overall cost (consumables and labor) of ddPCR was 2 times higher than rtPCR. 2,18 In terms of high-throughput detection, the ddPCR instrument has only 2 fluorescence channels, testing a maximum of 4 target genes; the rtPCR instrument has 5 fluorescence channels, testing a maximum of 5 target genes.…”

mentioning

confidence: 99%

Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus

Cao

Chen

et al. 2020

J VET Diagn Invest

Self Cite

View full text Add to dashboard Cite

Porcine epidemic diarrhea, a disease caused by porcine epidemic diarrhea virus (PEDV), results in large economic losses to the global swine industry. To manage this disease effectively, it is essential to detect PEDV early and accurately. We developed a sensitive and accurate droplet digital PCR (ddPCR) assay to detect PEDV. The optimal primer-to-probe concentration and melting temperature were identified as 300:200 nM and 59.2°C, respectively. The specificity of the ddPCR assay was confirmed by negative test results for common swine pathogens. The detection limit for the ddPCR was 0.26 copies/μL, which is a 5.7-fold increase in sensitivity compared to that of real-time PCR (rtPCR). Both ddPCR and rtPCR assays exhibited good linearity, although ddPCR provided higher sensitivity for clinical detection compared to that of rtPCR. Our ddPCR methodology provides a promising tool for evaluating the PEDV viral load when used for clinical testing, particularly for detecting samples with low-copy viral loads.

show abstract

mentioning

confidence: 99%

Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus

Cao

Chen

et al. 2020

J VET Diagn Invest

Self Cite

View full text Add to dashboard Cite

show abstract

“…On the basis of all the results of this research, we can state that the ddPCR is mainly suitable when excellent accuracy of results is required, while the qPCR is preferable if high detection efficiency is needed [ 24 ]. This statement is supported by the different SD values between these two methods ( Table 3 ).…”

Section: Resultsmentioning

confidence: 99%

Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese

Cutarelli

Fulgione

Fraulo

et al. 2021

Animals

View full text Add to dashboard Cite

Buffalo mozzarella cheese is one of the most appreciated traditional Italian products and it is certified as a Protected Designation of Origin (PDO) product under the European Commission Regulation No. 1151/2012. It is obtained exclusively from buffalo milk. If made from cow milk, or a mixture of buffalo and cow milk, buffalo mozzarella cheese does not qualify as a PDO product. In order to maximize their profits, some producers market buffalo mozzarella that also contains cow milk as a PDO product, thus defrauding consumers. New methods for revealing this fraud are therefore needed. One such method is the droplet digital Polymerase Chain Reaction (ddPCR). Thanks to its high precision and sensitivity, the ddPCR could prove an efficacious means for detecting the presence of cow milk in buffalo mozzarella cheese that is marketed as a PDO product. ddPCR has proved able to detect the DNA of cow and/or buffalo milk in 33 buffalo mozzarella cheeses labelled as PDO products, and experimental evidence could support its application in routine analyses.

show abstract

“…Despite its merits, the use of ddPCR still holds some disadvantages, such as higher operation cost, reagent costs, and availability, when compared to conventional methods [44,95]. Despite these limitations, the high accuracy and resolution of ddPCR has led it to be widely used in food sciences [31,45,[47][48][49]51,52,67,68,91,116,117] and disease and evolution studies [23,46,53,66,69,73,85,90,[92][93][94]99,101,[118][119][120][121][122][123]. These studies are continually developing the technology and creating an extensive collection and recording of the optimization process for different genes.…”

Section: Discussionmentioning

confidence: 99%

“…A study in the late 1980s by Clewley summarized the approaches to minimize contamination and erroneous issues of PCR by suggesting practical measures such as using well-characterized controls, improving lab techniques and practices, optimizing primer selection, and testing of plasmids [26]. Throughout the years, researchers also evaluated PCR for forensic applications [29], variant identification [30], and primer and temperature optimization [31,32], and PCR has been continually optimized for various applications [33][34][35]. What researchers expected as potential applications of the PCR technique have become more general nowadays.…”

Section: The Initial State Of Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

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)

Park

Rana

Sung

et al. 2021

Applied Microbiology

View full text Add to dashboard Cite

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.

show abstract

Species identification and quantification of silver pomfret using the droplet digital PCR assay

Cited by 24 publications

References 32 publications

Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus

Development of a droplet digital PCR for detection and quantification of porcine epidemic diarrhea virus

Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese

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)

Contact Info

Product

Resources

About