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摘要:Objective To develop a real-time PCR for the rapid detection of Vibrio parahaemolytichus in aquatic products. Methods A real-time PCR was developed for the rapid detection of Vibrio parahaemolytichus based on the primers and probe which were designed for the conservative domain of TDH gene. Results A real-time PCR for the rapid detection of Vibrio parahaemolytichus was developed.The quantitative detection limit of this method was less than 10cfu/PCR reaction for pure cultured broth, and 103cfu/g and 1 cfu/g for non-cultured and 4h-6h- cultured artificial contaminated aquatic products respectively. In the specificity test, among 24 reference/standard strains, only one strain of Vibrio parahaemolytichus was positive, while 23 strains of non- Vibrio parahaemolytichus were all negative. All of the coefficient of variation of intra-assay and inter-assay were less than 1%. Conclusion The real-time PCR is a repeatable, specific , sensitive and rapid method for the detection of Vibrio parahaemolytichus. It takes less than 8h to qualitatively or quantitatively detect Vibrio parahaemolytichus in aquatic products.

摘要:8-week-old female BALB/c mice were immunized with H4-AIV strain A/Duck/Yangzhou/185/2003 and the spleen cells were collected and fused with Sp2/0-Ag-14 myeloma cells.Hybridoma cells were screened by hemagglutination inhibition(HI)test and the positive clones were subjected to limited dilution for subcloning.4 mAbs against HA of H4-AIV were produced after triple subcloning and named as 2C11,6E7,4C8 and 5C5 re- spectively.All the 4 monoclonal antibodies were tested to be IgG2a subtype and the HI titers were between 1:212 and 1:215.Cross HI test and indirect immunofluorescence showed that the 4 mAbs were specific to H4-AIV as they reacted only with H4-AIV but not with NDV,IBV,EDSv-76 and other AIV subtypes.Virus neutralization test indicated that the 4 mAbs were able to inhibit the pathogenicity of H4-AIV in cell culture.The HI results re- vealed that mAbs 2C11,4C8,6E7 and 5C5 could inhibit 9,8,7 and 7 of 10 H4-AIV isolates respectively,while none of them could inhibit the A/Duck/Suzhou/1/2002 isolate,showing the antigenic difference among the HA pro- teins of the H4-AIV isolates.

摘要:A pair of PCR primers were designed according to the nucleocapsid protein sequences of Simian vires 5(SV5)in GenBank by Primer 5.0 software.The product is 265bp fragment.A RT-PCR was constructed method, which can be used to detect canine parainfluenza vires(CPIV).The specific experiment of PCR showed that the primers could only amplify the genome of CPIV.No any gene fragments had been amplified from the cells that were infected by canine distemper virus,canine parvovirus,canine adenovirus and rabies virus.The sensitivy ex- periment showed that the method could amplify the specific fragment from the soliquoid of spleen cell infected by CPIV at 10~(-5)dilution.It showed that this study provides an effective means for chnical rapid diagnosis of CPIV in CIRD.

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