Detection of sequence variation in PCR-amplified fragments of omp2 gene from three species of the family Chlamydiaceae using agarose gel electrophoresis containing bisbenzimide-PEG.
A simple technique providing a means for rapid genetic differentiation of chlamydial strains is described. The technique is based on a single-step sequence-specific separation of PCR-amplified DNA fragments by electrophoresis in an agarose gel containing a DNA ligand - bisbenzimide-PEG. A hypervariable region at the 5' end of the omp2 gene of Chlamydiaceae species encoding the 60-kDa cysteine-rich outer membrane protein was selected as a target for PCR. The appropriate fragments were amplified from strains of Chlamydia trachomatis, Chlamydophila pneumoniae, and Chlamydophila psittaci, and the PCR products originating from different species were electrophoretically separated in the presence of the DNA ligand. We therefore demonstrated that PCR with a single pair of primers followed by simple agarose gel electrophoresis with bisbenzimide-PEG can be applied to the differentiation of three members of the family Chlamydiaceae which are commonly recognized as human pathogens.
Demkin
V V
VV
Institute for Molecular Genetics RAS, Kurchatov Sq. 2, 123182, Moscow, Russia. vdemkin@img.ras.ru
Edelstein
M V
MV
Zimin
A L
AL
Edelstein
I A
IA
Suvorov
M M
MM
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D006690:Bisbenzimidazole / Q000031:analogs & derivatives; D002692:Chlamydia trachomatis / Q000145:classification / Q000235:genetics; D002693:Chlamydiaceae / Q000145:classification* / Q000235:genetics*; D016993:Chlamydophila pneumoniae / Q000145:classification / Q000235:genetics; D002691:Chlamydophila psittaci / Q000145:classification / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D014644:Genetic Variation; D006801:Humans; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C110412:H.A.-yellow; C063931:Omp2 protein, bacteria; D011092:Polyethylene Glycols; D006690:Bisbenzimidazole