Detection of plasmid DNA from all Chlamydia trachomatis serovars with a two-step polymerase chain reaction.
A polymerase chain reaction was used to amplify a 137-base-pair sequence of DNA from a Chlamydia trachomatis plasmid. Various parameters of the polymerase chain reaction were explored, and it was found that two short steps per reaction cycle were sufficient to achieve 10(12)-fold amplification in less than 1 h. By use of this procedure, 10(-18) g of a sequence of plasmid DNA, representing the amount of that sequence found in one C. trachomatis bacterium, was amplified to the point where it was clearly visible on an ethidium bromide-stained polyacrylamide gel under UV light. DNA from intact cells from each of the 15 serovars of C. trachomatis could also be amplified for visualization. With this procedure, the presence or absence of C. trachomatis DNA in a sample could be established in less than 1.5 h. The speed and extreme sensitivity of this detection procedure may make it a useful method for the detection of C. trachomatis, and similar techniques should be possible for any type of bacteria.
Welch
D
D
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202.
Lee
C H
CH
Larsen
S H
SH
D001483:Base Sequence; D002692:Chlamydia trachomatis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping
D004269:DNA, Bacterial