Cloning And Sequencing Of MOMP Gene Of Salpingitis Chlamydial In Hens And Detection With Dot Blot Hybridization

One strain of salpingitis chlamydial in hens isolated in Inner Mongolian was inoculated into SPF chicken embryos (5~7days old),Lj29 cells , Vero cells, BHK-21 cells respectively. Then it was cultured and observed. The results showed that this isolated strain of chlamydia only caused very few SPF chicken embryos to death and the situation of death had no law. The results also showed that the sequence of the degrees of these three cells sensitive to the strain was L929 cells > Vero cells > BHK-21 cells. According to the sequences of the conservative region of avain chlamydia MOMP genes reported by people abroad, one pair of primers were designed to amplify MOMP gene with PCR whose template was genome DNA of this strain, then it was sequenced. Lastly by using the technique of dot blot hybridization, the genome DNA of chlamydia was detected with the probe of MOMP gene labeled with Dig-11-dUTP by using the way of random primer.the results showed the degree of sensitivity of the probe was 10 pg and other pathogens could not be detected by this probe.By comparing the diagnostic ways of nucleotide probe and FC , the technique of nucleotide probe were proved to have high sensitivity and speci fi city.