title: [Multiplex PCR assay for detection of virulence genes in avian pathogenic Escherichia coli].
abstract: OBJECTIVE
  To efficiently study the virulence genes distribution of Avian pathogenic Escherichia coli (APEC), we developed four multiplex PCR to detect adhesin-associated genes, invasin and toxin-associated genes, serum resistance-associated genes and iron acquisition-associated genes.
  
  
  METHODS
  According to gene sequences published in GenBank, we designed and synthesized 18 specific primer pairs, which were used in the four multiplex PCR. Then, we determined the sensitivity of multiplex PCR using diluted bacterial or DNA templates. To verify the feasibility of these multiplex PCR, we determined the distribution of virulence genes in 100 APEC isolates using these multiplex PCR.
  
  
  RESULTS
  According to the results of PCR, we can conclude that each of the 18 genes was exactly and effectively amplified in the four multiplex PCR. The sensitivities of these four multiplex PCR were 10(3) Colony forming units (CFU), 10(3) CFU, 10(5) CFU, 10(5) CFU bacteria and 1ng, 1ng, 10ng and 10ng DNA, respectively. Furthermore, the results multiplex PCR for virulence genes distribution in 100 APEC were same as the single PCR.
  
  
  CONCLUSIONS
  These results suggest that multiplex PCR developed in this study could efficiently detect the virulence genes of APEC, which was a useful and rapid technique for epidemiological investigation.
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Meng
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Meng
authors.forename: Shaohui
authors.initials: S
authors.lastname: Wang
authors.forename: Xiangan
authors.initials: X
authors.lastname: Han
authors.forename: Yue
authors.initials: Y
authors.lastname: Han
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
authors.forename: Jianjun
authors.initials: J
authors.lastname: Dai
authors.forename: Shengqing
authors.initials: S
authors.lastname: Yu
mesh_terms: D000818:Animals; D002645:Chickens; D004372:Ducks; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000382:microbiology*; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D037521:Virulence Factors