Developing Low Density Gene Chips For Detection Of Sexually Transmitted Diseases Pathogens In Clinical Samples

Objective Low density gene chips for detection of sexually transmitted diseases (STDs)pathogens were developed by combined method of polymerase chain reaction(PCR) and reverse cross blot hybridization .Methods The species-specific probes for identification of five sexually transmitted diseases species were given homopolymer tails and then spotted onto a nylon membrane.the DNA fragment from each standard pathogens was amplified by PCR ,and the PCR products labeled with bio-16-dUTP during amplification were hybridized with species-specific probe on nylon memberane;the DNA fragment from clinical samples was amplified by PCR with three sets of primers and the PCR products labeled with bio-16-dUTP during amplification were also hybridized with five species-specific probes on nylon membrane.Results A 520bp DNA fragments were amplified from Neisseria gonococcus,Ureaplasma uralyticum , Chlamydia trachomatis ,a 350bp DNA fragments were amplified from Candida albicans, and a 167bp DNA fragments were amplified from HIV-1 env gene,the sensitivity could be improved to 20fg DNA.and PCR products were hybridized with the species-specific probe respectively on nylon membrane.Conclusion The result revealed that the method is sensitive rapid and specific. It is easy to interpret .The PCR products derived from different samples can be analyzed simultaneously, it is a very useful technology in clinical mycology.