2012Xumu shouyi xuebaoRequires access

Development of PCR Identification Method for Pasteurella multocida from Porcine

Lirui Li

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Abstract

The objective of this study was to develop a sensitive and specific PCR method for the detection of Pasteurella multocida from porcine.A pair of primers were designed according to the plpE genes of Pasteurella multocida published in GenBank.Then the specific PCR method on the basis of the plpE gene was developed and optimized for rapid detection of Pasteurella multocida.The sensitivity of the method were evaluated with the constructed recombinant plasmid of Pasteurella multocida and other strains.Finally the PCR method established was employed on the detection of 64 suspected samples collected from Sichuan province.Results showed the method based on the plpE gene can amplify the genome of Pasteurella multocida only,not for others and the detection limitation reached 5×101 copies.Thirty-six samples were infected with Pasteurella multocida and the positive rate reached 56.2%.These results indicate that the PCR method on the basis of the plpE gene is specific,repeatable and sensitive and can be used for the detection of Pasteurella multocida.

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What this paper is about

The objective of this study was to develop a sensitive and specific PCR method for the detection of Pasteurella multocida from porcine.A pair of primers were designed according to the plpE genes of Pasteurella multocida published in GenBank.Then the specific PCR method on the basis of the plpE gene was developed and optimized for rapid detection of Pasteurella multocida.The sensitivity of the method were evaluated with the constructed recombinant plasmid of Pasteurella multocida and other strains.Finally the PCR method established was employed on the detection of 64 suspected samples collected from Sichuan province.Results showed the method based on the plpE gene can amplify the genome of Pasteurella multocida only,not for others and the detection limitation reached 5×101 copies.Thirty-six samples were infected with Pasteurella multocida and the positive rate reached 56.2%.These results indicate that the PCR method on the basis of the plpE gene is specific,repeatable and sensitive and can be used for the detection of Pasteurella multocida.

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Available abstract

The objective of this study was to develop a sensitive and specific PCR method for the detection of Pasteurella multocida from porcine.A pair of primers were designed according to the plpE genes of Pasteurella multocida published in GenBank.Then the specific PCR method on the basis of the plpE gene was developed and optimized for rapid detection of Pasteurella multocida.The sensitivity of the method were evaluated with the constructed recombinant plasmid of Pasteurella multocida and other strains.Finally the PCR method established was employed on the detection of 64 suspected samples collected from Sichuan province.Results showed the method based on the plpE gene can amplify the genome of Pasteurella multocida only,not for others and the detection limitation reached 5×101 copies.Thirty-six samples were infected with Pasteurella multocida and the positive rate reached 56.2%.These results indicate that the PCR method on the basis of the plpE gene is specific,repeatable and sensitive and can be used for the detection of Pasteurella multocida.

Key concepts: Pasteurella multocida, Biology, GenBank, Pasteurella, Plasmid, Polymerase chain reaction, Gene, Microbiology

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