In-house and on-field validation of the multiplex PCR assay developed for authentication of three commercially important shrimp species

Wilwet, Lidiya; Jeyashakila, Robinson; Sivaraman, Balasubramanian; Nayak, Binaya Bhusan; Kumar, H. Sanath; Jaiswar, A. K.; Ratrey, Ved Prakash; Jeyasekaran, G.

doi:10.1016/j.lwt.2021.111701

Cited by 9 publications

(5 citation statements)

References 32 publications

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“…A number of different DNA-based procedures have been used widely for the identification and authentication of economically important fish species ( Carvalho, 2018 ; Böhme et al, 2019 ; Carvalho, Sampaio & Santos, 2020 ; Barbosa, Sampaio & Santos, 2021 ; Wilwet et al, 2021 ; Roungchun, Tabb & Hellberg, 2022 ). However, this is the first study to develop multiplex PCR protocols for the identification and authentication of the three principal catfish species of the genus Brachyplatystoma exploited commercially in Brazil ( B. rousseauxii, B. vaillantii , and B. filamentosum ), and which are the frequent targets of fraudulent substitution ( Carvalho, 2018 ; Carvalho, Sampaio & Santos, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

“…Despite their advantages, including their high yield and sensitivity, these modern methods require more complex laboratories, qualified personnel, and expensive reagents ( Creydt & Fischer, 2020 ; Pappalardo et al, 2022 ). Multiplex PCR, by contrast, is one of the traditional methods most used in forensic investigations, given its sensitivity, specificity, rapid throughput, relatively simple procedures, and low laboratory costs ( Veneza et al, 2017 ; Barbosa, Sampaio & Santos, 2021 ; Wilwet et al, 2021 ). Given this, the multiplex PCR protocols developed in the present study can be used routinely by fish processing plants to certify their products, and by government inspectors, to prevent fraudulent substitutions and mislabeling in the productive chain.…”

Section: Discussionmentioning

confidence: 99%

“…This technique involves the combination of different pairs of species-specific primers, which permit the unequivocal discrimination of the target taxa in the same reaction through the amplification of distinct banding patterns, which can be visualized in agarose gel ( Ali et al, 2012 ; Böhme et al, 2019 ). In recent years, several studies have developed multiplex PCR protocols for species identification, and have confirmed their accuracy, sensitivity, and robustness for the identification and authentication of fish samples ( Veneza et al, 2017 ; Kang, 2019 ; Barbosa et al, 2020 ; Barbosa, Sampaio & Santos, 2021 ; Wilwet et al, 2021 ). Among the markers used for the development of multiplex protocols for species identification purposes, mitochondrial DNA markers are known for their advantages such as the large number of copies per sample, the availability of sequences in public databases, as well as genomic regions with intraspecific conservation and interspecific polymorphism, which are necessary characteristics for the development of species-specific primers ( Hebert, Ratnasingham & DeWaard, 2003 ; Kang, 2019 ; Böhme et al, 2019 ).…”

Section: Introductionmentioning

confidence: 95%

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Development of rapid and cost-effective multiplex PCR assays to differentiate catfish of the genus Brachyplatystoma (Pimelodidae–Siluriformes) sold in Brazil

Freitas

Barbosa

Vale

et al. 2023

PeerJ

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The catfishes Brachyplatystoma filamentosum (Kumakuma), Brachyplatystoma vaillantii (Laulao catfish), and Brachyplatystoma rousseauxii (gilded catfish) are important fishery resources in Brazil, where they are sold both fresh and in the form of fillets or steaks. These species have morphological similarities, thus, they can be easily misidentified or substituted, especially after processed. Therefore, accurate, sensitive, and reliable methods are needed for the identification of these species to avoid commercial fraud. In the present study, we develop two multiplex PCR assays for the identification of the three catfish species. Each multiplex protocol combined three species-specific forward primers and a universal reverse primer to produce banding patterns able to discriminate the target species unequivocally. The length of the cytochrome C oxidase subunit I (COI) fragments was approximately 254 bp for B. rousseauxii, 405 bp for B. vaillantii, and 466 bp for B. filamentosum, while the control region (CR) assay produced fragments of approximately 290 bp for B. filamentosum, 451 bp for B. vaillantii, and 580 bp for B. rousseauxii. The protocols were sensitive enough to detect the target species at a DNA concentration of 1 ng/µL, with the exception of the CR of B. vaillantii, in which the fragment was only detectable at 10 ng/µL. Therefore, the multiplex assays developed in the present study were sensitive, accurate, efficient, rapid, and cost-effective for the unequivocal identification of the target species of Brachyplatystoma. They can be utilized by fish processing industries to certify their products, or by government agencies to authenticate products and prevent fraudulent commercial substitutions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

See 1 more Smart Citation

Development of rapid and cost-effective multiplex PCR assays to differentiate catfish of the genus Brachyplatystoma (Pimelodidae–Siluriformes) sold in Brazil

Freitas

Barbosa

Vale

et al. 2023

PeerJ

View full text Add to dashboard Cite

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“…The Asian regions—Southeast, East, and South Asia—face recurring food contamination issues due to dense populations, limited resources, and relatively less focus on food research. China, Malaysia, and India have encountered such incidents, prompting researchers’ increased development and validation of food safety protocols (Girish et al., 2020; Sheu et al., 2021; Wilwet et al., 2021; Xing et al., 2020). Mycotoxin contamination in various foods, from barley to processed meats, remains a significant concern (Lago et al., 2021; Rausch et al., 2021; Zadravec et al., 2020; Zhao et al., 2021).…”

Section: Food Authentication and Contamination In Asian Continentsmentioning

confidence: 99%

Food authentication, current issues, analytical techniques, and future challenges: A comprehensive review

Haider,

Iqbal,

Bhatti

et al. 2024

Comp Rev Food Sci Food Safe

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Food authentication and contamination are significant concerns, especially for consumers with unique nutritional, cultural, lifestyle, and religious needs. Food authenticity involves identifying food contamination for many purposes, such as adherence to religious beliefs, safeguarding health, and consuming sanitary and organic food products. This review article examines the issues related to food authentication and food fraud in recent periods. Furthermore, the development and innovations in analytical techniques employed to authenticate various food products are comprehensively focused. Food products derived from animals are susceptible to deceptive practices, which can undermine customer confidence and pose potential health hazards due to the transmission of diseases from animals to humans. Therefore, it is necessary to employ suitable and robust analytical techniques for complex and high‐risk animal‐derived goods, in which molecular biomarker‐based (genomics, proteomics, and metabolomics) techniques are covered. Various analytical methods have been employed to ascertain the geographical provenance of food items that exhibit rapid response times, low cost, nondestructiveness, and condensability.

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“…Improved traceability tools represent, in fact, an essential support for FBOs who, in any case, hold the responsibility of verifying the identity of their products for consumer protection in accordance with EU legislation [31]. In this regard, rapid identification methods have been promoted and developed in recent years for the most traded species, potentially applicable to self-monitoring by various operators, especially at the distribution level [80][81][82]. 3 presents the results of the comparison between the scientific names found in association with the products and the official list of accepted trade names in force during the sampling period [45], and the updated list [46].…”

Section: Mislabeling Rate and Products Originmentioning

confidence: 99%

An Authentication Survey on Retail Seafood Products Sold on the Bulgarian Market Underlines the Need for Upgrading the Traceability System

Tinacci

Stratev

Strateva

et al. 2023

Foods

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Economically motivated or accidental species substitutions lead to economic and potential health damage to consumers with a loss of confidence in the fishery supply chain. In the present study, a three–year survey on 199 retail seafood products sold on the Bulgarian market was addressed to assess: (1) product authenticity by molecular identification; (2) trade name compliance to the list of official trade names accepted in the territory; (3) adherence of the list in force to the market supply. DNA barcoding on mitochondrial and nuclear genes was applied for the identification of whitefish (WF), crustaceans (C) and mollusks (cephalopods—MC; gastropods—MG; bivalves—MB) except for Mytilus sp. products for which the analysis was conducted with a previously validated RFLP PCR protocol. Identification at the species level was obtained for 94.5% of the products. Failures in species allocation were reconducted due to low resolution and reliability or the absence of reference sequences. The study highlighted an overall mislabeling rate of 11%. WF showed the highest mislabeling rate (14%), followed by MB (12.5%), MC (10%) and C (7.9%). This evidence emphasized the use of DNA–based methods as tools for seafood authentication. The presence of non–compliant trade names and the ineffectiveness of the list to describe the market species varieties attested to the need to improve seafood labeling and traceability at the national level.

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In-house and on-field validation of the multiplex PCR assay developed for authentication of three commercially important shrimp species

Cited by 9 publications

References 32 publications

Development of rapid and cost-effective multiplex PCR assays to differentiate catfish of the genus Brachyplatystoma (Pimelodidae–Siluriformes) sold in Brazil

Development of rapid and cost-effective multiplex PCR assays to differentiate catfish of the genus Brachyplatystoma (Pimelodidae–Siluriformes) sold in Brazil

Food authentication, current issues, analytical techniques, and future challenges: A comprehensive review

An Authentication Survey on Retail Seafood Products Sold on the Bulgarian Market Underlines the Need for Upgrading the Traceability System

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