A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

Bai, Weibin; Xu, Wentao; Huang, Kunlun; Yang, Yuan; Cao, Sishuo; Luo, Yunbo

doi:10.1016/j.foodcont.2008.05.021

Cited by 49 publications

(34 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The adaptor is a 10-30nt random sequence which is rarely complementary to DNA template and primer. For example, when the forward primer of mice is modified with an adapter like CCTTCCTTCCTTCCCC CC as described by Bai et al [21], the resulted PCR product will be 146 bp, which is more reliable to discriminate from horse amplicon. Alternatively, PCR product can be run on polyacrylamide gel which has smaller mesh and better separation to similar-sized DNA fragments (as shown in Fig.…”

Section: Discussionmentioning

confidence: 99%

Species Authentication of Common Meat Based on PCR Analysis of the Mitochondrial COI Gene

Dai

Qiao

Yang

et al. 2015

Appl Biochem Biotechnol

View full text Add to dashboard Cite

Adulteration of meat products and costly animal-derived commodities with their inferior/cheaper counterparts is a grievous global problem. Species authentication is still technical challenging, especially to those deep processed products. The present study described the design of seven sets of species-specific primer based on a high heterozygous region of mitochondrial cytochrome c oxidase subunit I (COI) gene. These primers were proven to have high species specificity and no cross-reactions and unexpected products to different DNA source. Multiplex PCR assay was achieved for rapid and economical identification of four commonly consumed meats (pork, beef, chicken, and mutton). The conventional PCR assay was sensitive down to 0.001 ng of DNA template in the reactant. The developed method was also powerful in detecting as low as 0.1-mg adulterated pork (0.05 % in wt/wt) in an artificial counterfeited mutton. Validation test showed that the assay is specific, reproducible, and robust in commercial deep processed meats, leatherware, and feather commodities. This proposed method will be greatly beneficial to the consumers, food industry, leather, and feather commodity manufacture.

show abstract

Section: Discussionmentioning

confidence: 99%

Species Authentication of Common Meat Based on PCR Analysis of the Mitochondrial COI Gene

Dai

Qiao

Yang

et al. 2015

Appl Biochem Biotechnol

View full text Add to dashboard Cite

show abstract

“…This highlights a common problem with multiplex PCR reactions where sequences that are very abundant in the PCR matrix might The animal species marked in bold could be detected by the LCD Array kit and species specific PCR, but not by the CarnoCheck kit be over amplified and less abundant or trace sequences seemingly under represented. In the work by Bai et al 2009, the inherent complexity, low amplification efficiency, and unequal amplification efficiency on different templates were cited as major drawbacks of currently described multiplex PCR reactions, thus precluding their commercial application. Although the animal biochips here described exploit the principle of hybridization of the amplified fragments on specific probes bound on the arrays, thus heightening the sensitivity and specificity of this detection method, the procedure is nevertheless limited by the overall proficiency of the PCR amplification.…”

Section: Discussionmentioning

confidence: 99%

Biochip Technology for the Detection of Animal Species in Meat Products

Iwobi¹,

Huber²,

Hauner³

et al. 2010

Food Anal. Methods

View full text Add to dashboard Cite

The verification of declared components in meat products is an essential task of food control agencies worldwide. To date, the ELISA and species-specific polymerase chain reaction (PCR) are two commonly applied analytical tools employed by many authorized food control laboratories. These trusted methods however do not allow the simultaneous detection of all the animal species present in a meat sample. Additionally, detection of undeclared components resulting from inadvertent contamination or deliberate adulteration of the meat products requires additional processing of the samples, resulting in increased expenditure. The use of DNA biochip analysis that allows simultaneous processing of many meat products, while concomitantly generating results for the detection of all animal species present in the meat products is thus highly desirable. In this work, two commercially available animal chip detection systems (CarnoCheck Test Kit and MEAT species LCD Array) are compared in terms of sensitivity, robustness, reproducibility, and ease of handling. The two animal species differentiation biochip methods compared well in efficiency and could simultaneously detect from eight to 14 animal species in the meat products. Detection limits were found to be in the range of 0.1% to 0.5% in meat admixtures, with good reproducibility of results. More than 70 commercially available meat samples were analyzed in this work, with the results validated against traditional PCR methodology. Both biochip methods performed well and could be implemented for routine use in any food control agency.

show abstract

“…In cases where very small amounts of a meat species is present in the food matrix, the amplification of such sequences might be hampered by the presence of other meat species present in more abundance in the sample, leading to possible false negative results. Bai et al (2009) cited the inherent complexity, low amplification efficiency, and unequal amplification efficiency on different templates as major drawbacks of currently described multiplex PCR reactions, thus precluding their commercial application. The biochips here described nevertheless hold great promise in the parallel identification of meat species in food products or samples.…”

Section: Dna Chip Technology In Meat Species Differentiationmentioning

confidence: 99%