Quantitative Analysis of Pork and Chicken Products by Droplet Digital PCR

Cai, Yicun; Li, Xiang; Lv, Rong; Yang, Litao; Li, Jian; He, Yihua; Pan, Liangwen

doi:10.1155/2014/810209

Cited by 47 publications

(39 citation statements)

References 26 publications

(25 reference statements)

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“…In contrast, in another quantification study based on the ddPCR technique, which used a two-step conversion that increased the bias of quantification, the LOQ was 10 mg (10%) for pork, and the bias was approximately 17% [29]. In our study, the deviation of 10% (w/w) was approximately less than ±8.17%, and the LOQ was defined as 1%.…”

Section: Resultscontrasting

confidence: 55%

A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food

et al. 2017

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Meat adulteration is a worldwide concern. In this paper, a new droplet digital PCR (ddPCR) method was developed for the quantitative determination of the presence of chicken in sheep and goat meat products. Meanwhile, a constant (multiplication factor) was introduced to transform the ratio of copy numbers to the proportion of meats. The presented ddPCR method was also proved to be more accurate (showing bias of less than 9% in the range from 5% to 80%) than real-time PCR, which has been widely used in this determination. The method exhibited good repeatability and stability in different thermal treatments and at ultra-high pressure. The relative standard deviation (RSD) values of 5% chicken content was less than 5.4% for ultra-high pressure or heat treatment. Moreover, we confirmed that different parts of meat had no effect on quantification accuracy of the ddPCR method. In contrast to real-time PCR, we examined the performance of ddPCR as a more precise, sensitive and stable analytical strategy to overcome potential problems of discrepancies in amplification efficiency discrepancy and to obtain the copy numbers directly without standard curves. The method and strategy developed in this study can be applied to quantify the presence and to confirm the absence of adulterants not only to sheep but also to other kinds of meat and meat products.

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Section: Resultscontrasting

confidence: 55%

A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food

et al. 2017

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“…Fresh lean beef ( Bos taurus ), pork ( Sus scrofa ), chicken ( Gallus gallus ), and mutton ( Capra hircus ) were purchased from a local market in Shanghai, China. All the samples were minced, dried in a baking oven (UFE500AO, Memeert, Germany) at 80°C for 72 h, and ground into a superfine powder in liquid nitrogen using a bench-top 6850 Freezer/Mill set at 10 Hz for 8 min (SPEX SamplePrep, Metuchen, NJ) [ 16 ]. Mixed beef and pork powder samples of known proportion, ranging from 5%–to 95% beef/pork by mass (95 mg beef / 5 mg pork , 80 mg beef / 20 mg pork , 75 mg beef / 25 mg pork , 60 mg beef / 40 mg pork , 55 mg beef / 45 mg pork , 40 mg beef / 60 mg pork , 35 mg beef / 65 mg pork , 20 mg beef / 80 mg pork , 15 mg beef / 85 mg pork , 10 mg beef / 90 mg pork ), were prepared and used to assess the validity and sensitivity of the method.…”

Section: Methodsmentioning

confidence: 99%

Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR

Cai

et al. 2017

PLoS ONE

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Meat products often consist of meat from multiple animal species, and inaccurate food product adulteration and mislabeling can negatively affect consumers. Therefore, a cost-effective and reliable method for identification and quantification of animal species in meat products is required. In this study, we developed a duplex droplet digital PCR (dddPCR) detection and quantification system to simultaneously identify and quantify the source of meat in samples containing a mixture of beef (Bos taurus) and pork (Sus scrofa) in a single digital PCR reaction tube. Mixed meat samples of known composition were used to test the accuracy and applicability of this method. The limit of detection (LOD) and the limit of quantification (LOQ) of this detection and quantification system were also identified. We conclude that our dddPCR detection and quantification system is suitable for quality control and routine analyses of meat products.

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“…The intrinsic nature of ddPCR allows detection of rare nucleic acids in the presence of a high abundance of standard sequences (15). The applications of this technology have mainly been focused on clinical research and diagnosis, genetic modification, food product screening, and viral surveillance (17)(18)(19)(20)(21)(22)(23), while the potential of using ddPCR in environmental studies has yet to be explored.…”

mentioning

confidence: 99%

Comparison of Quantitative PCR and Droplet Digital PCR Multiplex Assays for Two Genera of Bloom-Forming Cyanobacteria, Cylindrospermopsis and Microcystis

Chen

Gin

2015

Appl Environ Microbiol

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The increasing occurrence of harmful cyanobacterial blooms, often linked to deteriorated water quality and adverse public health effects, has become a worldwide concern in recent decades. The use of molecular techniques such as real-time quantitative PCR (qPCR) has become increasingly popular in the detection and monitoring of harmful cyanobacterial species. Multiplex qPCR assays that quantify several toxigenic cyanobacterial species have been established previously; however, there is no molecular assay that detects several bloom-forming species simultaneously. Microcystis and Cylindrospermopsis are the two most commonly found genera and are known to be able to produce microcystin and cylindrospermopsin hepatotoxins. In this study, we designed primers and probes which enable quantification of these genera based on the RNA polymerase C1 gene for Cylindrospermopsis species and the c-phycocyanin beta subunit-like gene for Microcystis species. Duplex assays were developed for two molecular techniques-qPCR and droplet digital PCR (ddPCR). After optimization, both qPCR and ddPCR assays have high linearity and quantitative correlations for standards. Comparisons of the two techniques showed that qPCR has higher sensitivity, a wider linear dynamic range, and shorter analysis time and that it was more cost-effective, making it a suitable method for initial screening. However, the ddPCR approach has lower variability and was able to handle the PCR inhibition and competitive effects found in duplex assays, thus providing more precise and accurate analysis for bloom samples. C yanobacteria, also known as blue-green algae, constitute the most notorious phylum of phytoplankton capable of forming harmful blooms in freshwater aquatic ecosystems (1). Their presence in freshwater and brackish and coastal marine waters is of particular interest because of their massive accumulation and proliferation into nuisance blooms in nutrient-enriched water. These cyanobacterial blooms are the cause for a multitude of water quality concerns, due to their potential to produce secondary metabolites, some of which are toxins and compounds that compromise taste and odor. Cyanotoxins have been linked to human and animal illness and death and pose serious health hazards to communities which use surface waters and reservoirs as potable waters (2). Hence, the detection of cyanobacteria is crucial for reliable and prudent water management. The use of efficient detection methods in routine monitoring of waters is necessary to safeguard precious water resources.Microscopic counting combined with chemical detection of cyanotoxins in water samples is the conventional method to evaluate harmful cyanobacterial blooms (3). However, morphological identification is prone to limitations such as being time-consuming and unable to distinguish between toxic and nontoxic species and prone to misinterpretation when limited morphological differences are available (4, 5). In light of these discrepancies, DNAbased detection methods are becoming increasingly popular be...

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Quantitative Analysis of Pork and Chicken Products by Droplet Digital PCR

Cited by 47 publications

References 26 publications

A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food

A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food

Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR

Comparison of Quantitative PCR and Droplet Digital PCR Multiplex Assays for Two Genera of Bloom-Forming Cyanobacteria, Cylindrospermopsis and Microcystis

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