Skip to main content
SpringerLink
Log in

Loop-Mediated Isothermal Amplification Assays for the Rapid Identification of Duck-Derived Ingredients in Adulterated Meat

  • Published:
Food Analytical Methods Aims and scope Submit manuscript

Abstract

Meat adulteration is a growing concern in the marketplace today. To protect consumer rights and prevent unfair competition, it is essential to use an efficient assay to identify rapidly the species of meat being sold. In this context, a loop-mediated isothermal amplification (LAMP) assay coupled with a lateral flow dipstick (LFD) or hydroxynaphthol blue dye (HNB) was developed for the detection of duck genes in meat products. The LAMP-LFD and LAMP-HNB assays were performed at 65 °C for 30 min, with no cross-reactivity against four other species of meat. Sensitivity evaluation showed that the two assays can detect 3 pg of duck DNA per reaction, which is 10 times higher than that of the real-time quantitative polymerase chain reaction (qPCR). Through testing the experimental adulteration models, which were prepared by mixing duck meat with beef at different concentrations (0.01 to 10%), the detection limits of the two assays were confirmed as 0.1% duck meat. Combining the simple DNA extraction assay and the LAMP-LFD or LAMP-HNB assay, adulteration with only 1 mg duck meat can be identified within 40 min. The LAMP-LFD and LAMP-HNB assays are simple and highly sensitive, which provide valuable tools for the identification of duck gene from adulterated meat. The simple DNA extraction assay further enables our LAMP assays to be applicable in the field.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Abdulmawjood A, Grabowski N, Fohler S, Kittler S, Nagengast H, Klein G (2014) Development of loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive identification of ostrich meat. PLoS One 9(6):1–6

    Article  Google Scholar 

  • Ali ME, Hashim U, Mustafa S, Che Man YB (2012) Swine-specific PCR-RFLP assay targeting mitochondrial cytochrome B gene for semiquantitative detection of pork in commercial meat products. Food Anal Method 5(3):613–623

    Article  Google Scholar 

  • Ali ME, Rahman MM, Hamid SBA, Mustafa S, Bhassu S, Hashim U (2014) Canine-specific PCR assay targeting cytochrome b gene for the detection of dog meat adulteration in commercial frankfurters. Food Anal Method 7(1):234–241

    Article  Google Scholar 

  • Ayaz Y, Ayaz ND, Erol I (2006) Detection of species in meat and meat products using enzyme-linked immunosorbent assay. J Muscle Foods 17(2):214–220

    Article  Google Scholar 

  • Barakat H, El-Garhy HAS, Moustafa MMA (2014) Detection of pork adulteration in processed meat by species-specific PCR-QIAxcel procedure based on D-loop and cytb genes. Appl Microbiol Biotechnol 98(23):9805–9816

    Article  CAS  Google Scholar 

  • Ballin NZ, Vogensen FK, Karlsson AH (2009) Species determination - can we detect and quantify meat adulteration. Meat Sci 83(2):165–174

    Article  CAS  Google Scholar 

  • Boyaci IH, Uysal RS, Temiz T, Shendi EG, Yadegari RJ, Rishkan MM, Velioglu HM, Tamer U, Ozay DS, Vural H (2014) A rapid method for determination of the origin of meat and meat products based on the extracted fat spectra by using of Raman spectroscopy and chemometric method. Eur Food Res Technol 238(5):845–852

    Article  CAS  Google Scholar 

  • Cheng Y, Zhang MH, Hu K, Sun FD, TaoR GXJ, Luan FX (2014) Loop-mediated isothermal amplification for the event-specific detection of wheat B73-6-1. Food Anal Methods 7(2):500–505

    Article  Google Scholar 

  • Chandrasekar A, Raja A, Raj GD, Thangavelu A, Kumanan K (2015) Rapid detection of avian infectious bronchitis virus by reverse transcriptase-loop mediated isothermal amplification. Proc. Natl. Acad. Sci., India, sect. B Biol Sci 85(3): 815–820

  • D’Agostino M, Diez-Valcarce M, Robles S, Losilla-Garcia B, Cook N (2015) A loop-mediated isothermal amplification-based method for analysing animal feed for the presence of Salmonella. Food Anal Methods 8(10):2409–2416

    Article  Google Scholar 

  • Deng JR, Pei JJ, Gou HC, Ye ZD, Liu CC, Chen JD (2015) Rapid and simple detection of Japanese encephalitis virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick. J Virol Methods 213:98–105

    Article  CAS  Google Scholar 

  • Ding WC, Chen J, Shi YH, Lu XJ, Li MY (2010) Rapid and sensitive detection of infectious spleen and kidney necrosis virus by loop-mediated isothermal amplification combined with a lateral flow dipstick. Arch Virol 155(3):385–389

    Article  CAS  Google Scholar 

  • Duan YB, Ge CY, Zhang XK, Wang JX, Zhou MG (2014) A rapid detection method for the plant pathogen Sclerotinia sclerotiorum based on loop-mediated isothermal amplification (LAMP). Australasian Plant Pathol 43(1):61–66

    Article  CAS  Google Scholar 

  • Ebbinghaus P, Von SG, Krücken J (2012) Direct loop-mediated isothermal amplification from Plasmodium chabaudi infected blood samples: inability to discriminate genomic and cDNA sequences. Exp Parasitol 131(1):40–44

    Article  CAS  Google Scholar 

  • Fajardo V, González I, Rojas M, García T, Martín R (2010) A review of current PCR-based methodologies for the authentication of meats from game animal species. Trends Food Sci Technol 21(8):408–421

    Article  CAS  Google Scholar 

  • Francois P, Tangomo M, Hibbs J, Bonetti EJ, Boehme CC, Notomi T, Perkins MD, Schrenzel J (2011) Robustness of a loop-mediated isothermal amplification reaction for diagnostic applications. Pathogens and Disease 62(1):41–48

    CAS  Google Scholar 

  • Ghosh R, Nagavardhini A, Sengupta A, Sharma M (2015) Development of loop-mediated isothermal amplification (LAMP) assay for rapid detection of Fusarium oxysporum f. Sp. ciceris - wilt pathogen of chickpea. BMC Research Notes 8(1):40–50

    Article  Google Scholar 

  • Guan XY, GuoJC SP, Yang LT, Zhang DB (2010) Visual and rapid detection of two genetically modified soybean events using loop-mediated isothermal amplification method. Food Anal Methods 3(4):313–320

    Article  Google Scholar 

  • Kesmen Z, Yetiman AE, Sahin F, Yetim H (2012) Detection of chicken and Turkey meat in meat mixtures by using real-time PCR assays. Food Sci 77(2):167–173

    Article  Google Scholar 

  • Kokkinos PA, Ziros PG, Bellou M, Vantarakis A (2014) Loop-mediated isothermal amplification (LAMP) for the detection of salmonella in food. Food Anal Methods 7(2):512–526

    Article  Google Scholar 

  • Komatsu K, Maejima K, Fujita N, Netsu O, Tomomitsu T, Arie T, Teraoka T, Namba S (2015) A detection method based on reverse transcription loop-mediated isothermal amplification for a genetically heterogeneous Plantago asiatica mosaic virus. J Gen Plant Pathol 81(4):297–303

    Article  CAS  Google Scholar 

  • Luo JM, Xu ZQ, Nie K, Ding X, Guan L, Wang J, Xian YY, Wu XY, Ma XJ (2014) Visual detection of norovirus genogroup II by reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye. Food Environ Virol 6(3):196–201

    Article  CAS  Google Scholar 

  • Niu JH, Gao YR, Yin JM, Leng QY, Yang GS, Wang C, Ren Y (2015) Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of bacterial blight pathogen (Xanthomonas axonopodis pv. dieffenbachiae) in anthurium. Eur J Plant Pathol 142(4):801–813

    Article  CAS  Google Scholar 

  • Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop mediated isothermal amplification of DNA. Nucleic Acids Res 28(12):E63

    Article  CAS  Google Scholar 

  • Rohman A, Sismindari EY, Che Man YB (2011) Analysis of pork adulteration in beef meatball using Fourier transform infrared (FTIR) spectroscopy. Meat Sci 88(1):91–95

    Article  CAS  Google Scholar 

  • Stamoulisa P, Stamatisa C, Sarafidoua T, Mamuris Z (2010) Development and application of molecular markers for poultry meat identification in food chain. Food Control 21(7):1061–1065

    Article  Google Scholar 

  • Sun M, Gao HW, Xiao XZ, Chen JX, Liu CX, Feng LP (2015) A novel loop-mediated isothermal amplification method for detection of the carrot materials in foods. Eur Food Res Technol 241(2):295–302

    Article  CAS  Google Scholar 

  • Thongkao K, Longyant S, Silprasit K, Sithigorngul P, Chaivisuthangkura P (2015) Rapid and sensitive detection of Vibrio harveyi by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to vhhP2 gene. Aquac Res 46(5):1122–1131

    Article  CAS  Google Scholar 

  • Uematsu H, Inoue Y, Ohto Y (2015) Detection of Pantoea stewartii from sweet corn leaves by loop-mediated isothermal amplification (LAMP). J Gen Plant Pathol 81(3):173–179

    Article  CAS  Google Scholar 

  • Wang F, Jiang L, Ge BL (2012) Loop-mediated isothermal amplification assays for detecting shigatoxin-producing Escherichia coli in ground beef and human stools. J Clin Microbiol 50(1):91–97

    Article  Google Scholar 

  • Wang W, Zhu YP, Chen Y, Xu XL, Zhou GH (2015) Rapid visual detection of eight meat species using optical thin-film biosensor chips. J AOAC Int 98(2):410–414

    Article  CAS  Google Scholar 

  • Zahradnik C, Martzy R, Mach RL, Krska R, Farnleitner AH, Brunner K (2015) Loop-mediated isothermal amplification (LAMP) for the detection of horse meat in meat and processed meat products. Food AnalN 8(6):1576–1581

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Centre for Disease Control and Prevention, Hangzhou, Zhejiang Province, People’s Republic of China

    Ya Shi, Yan Feng, Changping Xu, Zhouheng Xu & Yiyu Lu

  2. College of Life Science, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China

    Ya Shi

  3. College of Medical Technology, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China

    Dongqing Cheng

Authors
  1. Ya Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Changping Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhouheng Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dongqing Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yiyu Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Dongqing Cheng or Yiyu Lu.

Ethics declarations

Funding

This study was financially supported by the fund of Key Medical Subjects Construction Project of Zhejiang Province (XKQ-009-003), and the analytical test technology project of Zhejiang Province (2016C37006).

Conflict of Interest

Ya Shi declares that he has no conflict of interest. Yan Feng declares that he has no conflict of interest. Changping Xu declares that he has no conflict of interest. Zhouheng Xu declares that he has no conflict of interest. Dongqing Cheng declares that he has no conflict of interest. Yiyu Lu declares that he has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shi, Y., Feng, Y., Xu, C. et al. Loop-Mediated Isothermal Amplification Assays for the Rapid Identification of Duck-Derived Ingredients in Adulterated Meat. Food Anal. Methods 10, 2325–2331 (2017). https://doi.org/10.1007/s12161-016-0767-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12161-016-0767-0

Keywords

Search

Navigation