Towards a quantitative application of real-time PCR technique for fish DNA detection in feedstuffs

Benedetto, Alessandro; Abete, Maria Cesarina; Squadrone, Stefania

doi:10.1016/j.foodchem.2010.11.131

Cited by 27 publications

(18 citation statements)

References 23 publications

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“…Compared to other PCR systems published, the developed method is less sensitive. Benedetto et al [9] described a real-time PCR method to detect fish DNA in feedstuff based on the mitochondrial 12S rRNA gene. With the primer-probe system generated, a limit of detection of 0.2 pg fish DNA diluted in plant DNA could be reached.…”

Section: Sensitivitymentioning

confidence: 99%

“…For detection of fish allergens in food, enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) technology are used most widely [7][8][9][10][11]. The majority of ELISA systems are based on the detection of the major fish allergenic protein, parvalbumin [8].…”

Section: Introductionmentioning

confidence: 99%

“…It refers to the Combined Nomenclature listing bony and cartilaginous fish. Most of the detection systems for fish available at present are specific for only a limited number of fish species, for specific allergens or use broadly reactive mitochondrial genes as target [9][10][11][12][13]. In this work, a real-time PCR system for the detection of all relevant species of fish which may be present in food products was developed.…”

Section: Introductionmentioning

confidence: 99%

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Development of a real-time PCR system for the detection of the potential allergen fish in food

Tetzlaff

Mäde

2016

Eur Food Res Technol

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Section: Sensitivitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development of a real-time PCR system for the detection of the potential allergen fish in food

Tetzlaff

Mäde

2016

Eur Food Res Technol

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“…The amplification of a specific DNA sequence by PCR provides a rapid, sensitive and specific method for the detection of animal tissues in food and feed [35]. However, many PCR-based methods cannot be used for the detection of MBM, since the high temperatures used in the standard rendering process cause DNA fragmentation, leading to difficulty in obtaining reliable results [36].…”

Section: Spiked Feeds Concentration Of Mbm (%) W/w Addedmentioning

confidence: 99%

“…For the analyses involving different vertebrate species, two mitochondrial genes were used: 12S rRNA caprine speciespecific and cytochrome b for bovine, ovine, porcine and avian material (chicken). For the endogenous control the tRNA leucine gene was analyzed, which is a sequence present in the chloroplast genome [35].…”

Section: Spiked Feeds Concentration Of Mbm (%) W/w Addedmentioning

confidence: 99%

Detection of Animal Products in Ruminant Feeds by Microscopy and Real Time PCR

Golinelli¹,

Silva²

2015

J Veterinar Sci Technol

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This study evaluated the performance of the TaqManTM real-time PCR assay to detection of DNA from not allowed-animal derivatives in reference feeds samples. The results of qPCR were compared with the microscopy, only method validated to control the presence of animal proteins, according the European Communities. The qPCR tests targeting 12S rRNA from cows, sheep, porcine and chickens and cytochrome b region from caprine in feeds were able to detect half the amount (0.0125% w/w) of meat-and-bone meal (MBM) that could be detected by microscopy in samples spiked with MBM. Although cross-contamination in feeds and food processing plants is an unexceptional problem, the presence of traces of prohibited animal products in feedstuffs is an alert to potential impact on herd and human health, because it has been associated with the transmission of transmissible spongiform encephalopathy (TSE). These results indicate that a combination of qPCR tests and microscopic analysis could be used to ensure the safety of feedstuffs, allowing the identification of the animal species of the derivative and even the kind of tissue added to the feed, providing useful information for sanitation inspection authorities in this country.

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A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences

Eischeid

2018

J Sci Food Agric

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BACKGROUND: Fish are one of eight major allergens defined in the US Food Allergen Labeling and Consumer Protection Act, and cod and pollock are two of the major fish allergens. This paper describes development and validation of a method to detect cod and pollock in complex food matrices using real-time polymerase chain reaction (PCR). Mitochondrial cytochrome oxidase I (COI) sequences obtained through DNA barcoding were used to design a single set of primers and probe which detected three species in the genus Gadus: Atlantic cod, Pacific cod, and walleye pollock. RESULTS: Cod spiked into three different food matrices (cooking oil, clam chowder, and hushpuppy mix) yielded high linearity, dynamic range spanning six orders of magnitude, and lower limits of detection at 1-10 ppm (ppm; mg kg −1 ). Frying had an adverse effect on the lower limit of detection, but not on linearity.CONCLUSIONS: This work shows that COI DNA barcoding sequences can be used to effectively design real-time PCR assays for detection of food allergens in complex matrices. While full-length DNA barcodes distinguish individual species, the PCR assay designed here detected three different species. This is likely because real-time PCR assays are tolerant to basepair mismatches and do not utilize the full length of the DNA barcode. Published 2018. This article is a U.S. Government work and is in the public domain in the USA.

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Towards a quantitative application of real-time PCR technique for fish DNA detection in feedstuffs

Cited by 27 publications

References 23 publications

Development of a real-time PCR system for the detection of the potential allergen fish in food

Development of a real-time PCR system for the detection of the potential allergen fish in food

Detection of Animal Products in Ruminant Feeds by Microscopy and Real Time PCR

A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences

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