multiplex pcr (3)

Chinese researchers have developed a multiplex PCR assay using universal primers based on mitochondrial DNA to identify 8 species (dog, chicken, cattle, pig, horse, donkey, fox, and rabbit) simultaneously in meat products. The assay was tested on 103 commercial meat products from the Chinese market, which demonstrated its effectiveness and applicability.

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Researchers have a developed a multiplex PCR (polymerase chain reaction) assay to identify beef, pork, horse and poultry (chicken, turkey) and determine these species quantitatively in meat products. The qualitative assay uses a mitochondrial cytochrome b gene marker. The quantitative assay uses singlecopy markers from chromosomal genes (cyclic-GMP-phosphodiesterase gene for cattle, beta-actin gene for pig, interleukin-2 gene for chicken), and the normaliser is from the myostatin gene for mammals and poultry. The reliability of both methods was confirmed by analysing of mixed samples prepared with or without heat treatment. The assay was tested with 14 meat products from the Czech retail market with two having undeclared species and another 4 products giving an incorrect quantitative declaration. 

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A lab-on-a-chip-based multiplex polymerase chain reaction (PCR) assay for the authentication of five non-halal meat species in foods is described. Using species-specific primers, 172, 163, 141, 129 and 108-bp sites of mitochondrial ND5, ATPase 6 andcytochrome b genes were amplified to detect cat, dog, pig, monkey and rat species under complex matrices. Species-specificity was authenticated against 20 different species with the potential to be used in food. The assay was optimised under the backgrounds of various commercial meat products and validated for the analysis of meatballs, burgers and frankfurters, which are popular fast food items across the globe. The assay was tested to detect 0.1% suspected meats under commercial backgrounds of marketed foods.

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