Development of a direct loop-mediated isothermal amplification (LAMP) assay for rapid and simple on-site detection of chicken in processed meat products

Abstract We developed a direct loop-mediated isothermal amplification (LAMP) assay targeting the mitochondrial 16S rRNA gene to detect chicken in processed meat products. This rapid on-site detection method was developed using a portable fluorescence device and the LAMP assay with direct amplification. Chicken-specific LAMP primer set was designed and its specificity confirmed against 24 other animal species. The limit of detection (LOD) of the chicken-specific LAMP assay for chicken DNA was 10 fg. To evaluate the sensitivity of the direct LAMP assay, different percentages of chicken meat prepared with pork or beef were prepared. Heat- and pressure-treated meat mixtures were used to assess the applicability of this assay. LODs of the direct LAMP assay for the chicken/pork and chicken/beef mixtures were 0.1% for raw meat and 1% for heat- and pressure-treated meat. The direct LAMP assay was applied to 19 commercially available processed meat products. This assay successfully amplified the target sequence and identified the presence of chicken meat in processed meat products within approximately 30 min. This direct LAMP assay is a rapid and simple method for on-site detection of chicken meat in commercial products to monitor the authentication of chicken in meat products.

[1]  Mi-Ju Kim,et al.  Direct duplex real-time loop mediated isothermal amplification assay for the simultaneous detection of cow and goat species origin of milk and yogurt products for field use. , 2018, Food chemistry.

[2]  C. Hitchcock,et al.  Methodology for meat species identification: A review. , 1985, Meat science.

[3]  K. Holmstrøm,et al.  Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions. , 1992, International journal of food microbiology.

[4]  T. Notomi,et al.  Loop-mediated isothermal amplification of DNA. , 2000, Nucleic acids research.

[5]  C. Cammà,et al.  Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures , 2012 .

[6]  N. Grabowski,et al.  Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid and Sensitive Identification of Ostrich Meat , 2014, PloS one.

[7]  T. Notomi,et al.  Accelerated reaction by loop-mediated isothermal amplification using loop primers. , 2002, Molecular and cellular probes.

[8]  Y. Hagihara,et al.  On-site identification of meat species in processed foods by a rapid real-time polymerase chain reaction system. , 2017, Meat science.

[9]  T. García,et al.  Authentication of meat and commercial meat products from common pigeon (Columba livia) woodpigeon (Columba palumbus) and stock pigeon (Columba oenas) using a TaqMan® real-time PCR assay , 2012 .

[10]  R. Whittaker,et al.  An enzyme‐linked immunosorbent assay for species identification of raw meat , 1983 .

[11]  Ae-Ri Cho,et al.  Meat Species Identification using Loop-mediated Isothermal Amplification Assay Targeting Species-specific Mitochondrial DNA , 2014, Korean journal for food science of animal resources.

[12]  J. Spearow,et al.  The effects of curing and cooking on the detection of species origin of meat products by competitive and indirect ELISA techniques. , 1987, Meat science.

[13]  Y. Kumar,et al.  Loop-Mediated Isothermal Amplification (LAMP): A Rapid and Sensitive Tool for Quality Assessment of Meat Products. , 2017, Comprehensive reviews in food science and food safety.

[14]  Adrian Linacre,et al.  Forensic animal DNA analysis using economical two-step direct PCR , 2014, Forensic Science, Medicine, and Pathology.

[15]  M. Ahmed,et al.  Meat species identification using DNA-redox electrostatic interactions and non-specific adsorption on graphene biochips , 2016 .

[16]  M. Shimokomaki,et al.  Hamburger meat identification by dot-ELISA. , 2000, Meat science.

[17]  Noor Faizah Mohd-Naim,et al.  From market to food plate: Current trusted technology and innovations in halal food analysis , 2016 .

[18]  Enrique Sentandreu,et al.  Authenticity of meat products: Tools against fraud , 2014 .

[19]  Masato Saito,et al.  Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor , 2010 .

[20]  A. Piestrzynska-Kajtoch,et al.  The species identification of bovine, porcine, ovine and chicken components in animal meals, feeds and their ingredients, based on COX I analysis and ribosomal DNA sequences , 2013 .

[21]  J. Ongerth,et al.  Development and Preliminary Evaluation of a Loop-Mediated Isothermal Amplification Procedure for Sensitive Detection of Cryptosporidium Oocysts in Fecal and Water Samples , 2007, Applied and Environmental Microbiology.

[22]  S. Garrett,et al.  Detection of meat species using TaqMan real-time PCR assays. , 2004, Meat science.

[23]  Deepak Sharma,et al.  Identification of Species Origin of Meat and Meat Products on the DNA Basis: A Review , 2015, Critical reviews in food science and nutrition.

[24]  M. T. Bottero,et al.  Animal species identification in food products: evolution of biomolecular methods. , 2011, Veterinary journal.

[25]  D. Cawthorn,et al.  What is the role and contribution of meat from wildlife in providing high quality protein for consumption , 2012 .

[26]  Phuvadol Thanakiatkrai,et al.  Direct-multiplex PCR assay for meat species identification in food products. , 2014, Food chemistry.

[27]  Shin-Young Lee,et al.  Development of a rapid on-site detection method for pork in processed meat products using real-time loop-mediated isothermal amplification , 2016 .

[28]  Isabel Mafra,et al.  Food authentication by PCR-based methods , 2008 .