A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species |
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| Authors: | Weibin Bai Wentao Xu Kunlun Huang Yanfang Yuan Sishuo Cao Yunbo Luo |
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| Affiliation: | 1. Laboratory of food safety and molecular biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;2. Supervision and Testing Center of Agricultural Products Quality, Ministry of Agriculture, Beijing 100083, China;1. National Research Centre on Meat, Chengicherla, Hyderabad 500092, India;2. Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences, Hyderabad 500030, India;3. Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences, Tirupati 517501, India;4. Proteomics Lab, National Centre for Cell Science, Pune 411007, India;1. Nanotechnology and Catalysis Research Centre (NanoCat), University of Malaya, Kuala Lumpur 50603, Malaysia;2. Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, Kuala Lumpur 50603, Malaysia;1. Food Safety and Hygiene Division, Department of Environmental Health Engineering, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran;2. Food and Drug Control Reference Laboratories Center, Food and Drug Organization, Ministry of Health & Medical Education, Tehran, Iran;3. Reference Health Laboratories Research Center, Ministry of Health & Medical Education, Tehran, Iran |
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| Abstract: | A novel common primer multiplex PCR (CP-M-PCR) was applied to detect four kinds of meats (chicken, cattle, pig and horse) as raw materials. A common adapter was designed in the 5′-end of species-specific reverse primers which matched with the species-specific DNA sequences for each species and also used as the common primer (CP). CP-M-PCR primers were designed to uncover different length fragments of 239, 292, 412, and 451 bp from chicken, cattle, pig and horse meats, respectively. The bands of specific DNA fragments amplified by CP-M-PCR method still appeared until the concentration of species-specific primers diluted to 0.015 pmol and primer sensitivity was increased by 100 times compared with conventional multiplex PCR without CP. CP-M-PCR detection limit of the DNA samples was 0.1 ng (36.4 copies) for single kind of meat as well as four kinds of meats. CP-M-PCR method simplified the PCR reaction system and conquered the disparate amplified efficiency from different primers. The CP-M-PCR method could be widely applied in practical detection for simultaneous identification of other meat species and their products. |
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