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

Cai, Yicun; He, Yihua; Lv, Rong; Hong-chao, Chen; Wang, Qiang; Pan, Liangwen

doi:10.1371/journal.pone.0181949

Cited by 69 publications

(38 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results support previous findings, as PCI method has been used in many DNA isolation studies in this field [3,7,12,15]. Compared to PCI method, isolation kit method showed relatively low DNA yield, albeit having good DNA quality.…”

Section: Results and Discussion Conventional Pci Methods Showed Reliablsupporting

confidence: 90%

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Yahya

Firmansyah

Arlisyah³

et al. 2017

JELS

View full text Add to dashboard Cite

Detection of porcine contamination in food material by employing PCR techniques is integral in halal food confirmation. However, PCR is both costly and laborious, particularly in DNA isolation method. This study explores several different methods in DNA extraction for PCR amplification in bovine and porcine raw and boiled meat samples. Four methods for DNA extraction (conventional PCI method, DNA isolation kit, alkaline-based method, and a DNA lysis buffer-only from the same kit) was employed followed by PCR using primers from previous studies and compared for DNA quality and quantity (in six replicates) and PCR amplification on the best three DNA samples. This study shows that in all samples, the conventional method had the best DNA yield based on nanodrop measurement, followed by an alkali-based method, buffer-only method, and DNA isolation kit. Each method except lysis-buffer only had at least one sample with good DNA quality. Conventional and isolation kit showed reliable positive PCR detection for all porcine and bovine samples (92% positive). Using the alkaline-lysis method, DNA was amplified reliably on boiled meat samples (83% positive). Lysis-buffer-only method did not show consistent PCR amplification on the samples used (50% positive). The conclusion was that conventional PCI method and DNA isolation kit showed high reliability in PCR amplification of bovine and porcine meats, both raw and boiled. While high DNA yield was obtained using the alkaline-lysis method, PCR amplification was only successful on boiled samples. Lysis-buffer only method yielded in poor DNA quality and was not able to result in reliable DNA amplification.

show abstract

Section: Results and Discussion Conventional Pci Methods Showed Reliablsupporting

confidence: 90%

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Yahya

Firmansyah

Arlisyah³

et al. 2017

JELS

View full text Add to dashboard Cite

show abstract

“…It is important to note that Poland is one of the leading European exporters of game: in 2014, 3363 tonnes of wild boar meat was exported, representing over 30% of all exported game meat [23]. It was also found that meat products are often produced by combining meat from different animal species, which significantly increases the risk of infection [1,6].…”

Section: Discussionmentioning

confidence: 99%

The Seroprevalence of Toxoplasma gondii in Wild Boars from Three Voivodeships in Poland, MAT Analyses

et al. 2020

View full text Add to dashboard Cite

Purpose The European wild boar (Sus scrofa) is a popular game animal species. Its meat, however, can represent a reservoir of dangerous foodborne diseases and can play an important role in the transmission of many pathogens, including Toxoplasma gondii, in humans and animals worldwide. The aim of the present study was to determine the presence of antibodies to T. gondii in the serum of hunted wild boars in Poland. Methods Using the commercial direct agglutination test, 398 serum samples collected during the hunting season 2009/2010 were tested for the presence of T. gondii antibodies, and the titre of 40 was considered indicative of T. gondii infection in analysed samples. Results It was found that nationwide, 37.7% were seropositive to T. gondii, although seroprevalence varied from 11.6 to 50% depending on the Voivodeship. Significant differences were observed between the Great Poland and Lubusz Voivodeships and between Great Poland and Warmian-Masurian. Conclusion Serological test indicated widespread exposure to T. gondii by wild boar; therefore, consumption of raw or undercooked game meat of infected animals can carry a significant risk of T. gondii infection.

show abstract

“…Rising consumer demand underscores the need for the development of more swift and reliable methods to identify species in food commodities such as detecting pork in food. On this note, instead of protein, a DNA analysis would be preferable to identify species due to the nature of protein being easily denaturised while processing [1][2][3][4].…”

Section: Introductionmentioning

confidence: 99%

Electrochemical DNA biosensor for detection of pork (Sus scrof) using screen printed carbon-reduced graphene oxide electrode

Hartati¹,

Setiawati²,

Sofyatin³

et al. 2020

ScienceAsia

View full text Add to dashboard Cite

The identification of pork in foodstuff is critical regarding the counterfeiting of meat and kosherness, which is a particular concern for certain religions. In this study, we developed an electrochemical detection method of pork DNA without the use of DNA amplification by using screen printed carbon-reduced graphene oxide (SPC-RGO) electrode. The probe DNA of CytB gene of S. scrofa mtDNA was immobilized on the SPC-RGO surface by passive adsorption. Differential pulse voltammetry (DPV) was used to characterise the probe-target DNA hybridisation based on the target's guanine oxidation signal. The Placket-Burman and Box Behnken designs were used to select the factors that influence the hybridisation of probe-target DNA and to optimise each parameter. The following findings regarding the several factors that influence the hybridisation process and optimum condition were obtained: 5.0 µg/ml of probe DNA, 6.0 min of immobilisation time of probe DNA, 20.0 min of probe-target hybridisation time, a scan rate at 0.5 V/s, the pulse amplitude at 50.0 mV, and the washing time of the electrode being as long as 40 s. The limit of detection was obtained at 1.76 µg/ml for the linear range of 0-10.0 µg/ml target DNA while the relative standard deviation (RSD) was 2.25%. The DNA biosensor was tested on the isolated DNA samples from pork, chicken and beef while the voltammetry response reveals that it can distinguish the samples. These results indicate that the proposed electrochemical DNA biosensor has the potential to develop the detection method of pork content in the food samples.

show abstract

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

Cited by 69 publications

References 23 publications

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

Comparison of DNA Extraction Methods Between Conventional, Kit, Alkali and Buffer-Only for PCR Amplification on Raw and Boiled Bovine and Porcine Meat

The Seroprevalence of Toxoplasma gondii in Wild Boars from Three Voivodeships in Poland, MAT Analyses

Electrochemical DNA biosensor for detection of pork (Sus scrof) using screen printed carbon-reduced graphene oxide electrode

Contact Info

Product

Resources

About