Development and Application of a PCR Approach for Detection of Bovis, Sheep, Pig, and Chicken Derived Materials in Feedstuff |
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Authors: | LUO Jia-qin WANG Jia-qi BU Deng-pan LI Dan WANG Li WEI Hong-yang ZHOU Ling-yun |
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Affiliation: | 1. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences/Milk and Dairy Quality Supervision and Inspection Evaluation Center, Ministry of Agriculture, Beijing 100193, P.R.China;School of Animal Scienc 2. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences/Milk and Dairy Quality Supervision and Inspection Evaluation Center, Ministry of Agriculture, Beijing 100193, P.R.China |
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Abstract: | A PCR method for detection of bovis, sheep, pig, and chicken derived materials in feedstuff was established, and the existing method was improved according to the research on general primer and species-specific primers. First, general primer designed according to 16S rRNA gene sequence of bovis, sheep, pig, chicken, fish, and horse mtDNA was used for primary detection of animal derived materials in feedstuff. Species-specific primers designed according to conserved sequence of mtDNA of bovis, sheep, pig, and chicken were used for amplification of a 271, 274, 149, and 266 bp fragment, respectively. Further confirmation of the detection result was then carried out. PCR method for detection of animal derived materials in unknown feedstuff was developed by using general primer, relevant PCR system, and PCR condition. Also a PCR method for detection of each species (bovines, sheep, pig, and chicken) was designed by using our species-specific primers. High sensitivity and specificity of our method were confirmed with a minimum detection level of 0.1%. Method for detection of animal derived materials in this research is not only cheap and easy for operation but also precise and reliable results can be obtained. It could be one of the effective methods for the detection of animal derived materials in feedstuff. |
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Keywords: | animal feedstuff sensitivity and specificity PCR primer |
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