Improved PCR sensitivity for direct genotyping of Chlamydia trachomatis serovars by using a nested PCR.
Successful amplification of omp1 DNA by PCR is crucial in the genotyping of Chlamydia trachomatis when directly performed with clinical samples (J. Lan, J. M. M. Walboomers, R. Roosendaal, G. J. van Doornum, D. M. McLaren, C. J. L. M. Meijer, and A. J. C. van den Brule, J. Clin. Microbiol. 31:1060-1065, 1993). Several primers flanking the four variable domains of the omp1 gene were selected and tested for sensitivity in several nested PCRs with serial dilutions of serovar G. The optimal sensitivity obtained was 0.1 to 0.01 inclusion-forming units, similar to that obtained in the C. trachomatis plasmid PCR. With this approach, any C. trachomatis PCR-positive sample can be typed.
Lan
J
J
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Ossewaarde
J M
JM
Walboomers
J M
JM
Meijer
C J
CJ
van den Brule
A J
AJ
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D002690:Chlamydia Infections / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D018272:Porins; D012680:Sensitivity and Specificity; D012703:Serotyping
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D018272:Porins; C071677:omp1 protein, Chlamydia trachomatis