Investigation The AR Epidemiology In Guangxi And Cloning And Sequence Analysis Of The Swine OMPH Gene Of Pasteurlla Multocida

In this study,we respectively designed a couple of special primer(Bl/B2 and TOXA3/TOXA4)according to the Flagellin gene of Bb of and toxA of T~+Pm which are used to construct the detected system of multiple polymerase chain reaction on AR.The resules demonstrated this multiplicitas PCR was very specific and sensitive.353nose swabs from 10, different countries in Guangxi were detected by multiple PCR which have been constructed .The results of detected rate of T~+Pm PCR and Bb PCR showed that the positive number was 110,59 respectively. T~+Pm and Bb were not only detected from the swines which had the AR clinical symptom, also dissociated from non- clinical symptomatic swine. The data suggested T~+Pm and Bb were prevalent in the hoggerys of Guangxi,and mainly show subclinical infection.The distribution of AR in Guangxi as follows: AR prevalented seriously in the southeast of Guigang( Yulin, Luchuan,Bobai); Nanning and its periphery region ( Wuming, Fusui,Congzuo); the north of Guangxi (Liuzhou); the south of Guangxi(Hepu).The results can duce to take step to purification AR in the hoggerys as soon as possible. So it is necessary to construct a multiple PCR for the epidemiological survey of T~+Pm and Bb of AR in Guangxi.,and issignificance to the development of hog industry in Guangxi.OMPH is the major outer membrane protein of Pm, OMPH plays an important role in infection and pathogenesis. We amplified the OMPH gene of eight Pm isolates from Guangxi by polymerase chain reaction(PCR) with primers OMPH3/ OMPH4 . The eight OMPH genes were cloned and then sequenced. We compared the nucleotide sequence of OMPH gene ORF , their deduced amino acid sequences and analysed phylogenetic tree with those of OMPH gene nucleotide sequence published on GeneBank attempe to know some biological characteristics of Pm OMPH gene in Guangxi on the molecule level.. The results indicated all Pm isolates from Guangxi could be divided into two groups as I and II. Seven isolates GXHP,GXCZ,GXLZ,GXFS,GXWM,GXNN, GXGG belong to group I . GXYL belongs to group II. The mutations of OMPH gene focus 45,46,82,83,85,91,101,182,188,218,223,258,272,322,324 and 326amino acid,all mutations of amino acid should specificity of the group . The nucleotide homologies and the homologies of deduced amino acid of the eight isolates from Guangxi were 90.9-100.0%.The homology was very high .We can understand the relations between Guangxi and other strains in genetics and evolutions rule by the results. To study the feasibility for application vaccine and provide good basis on development of more available new vaccine for Guangxi.