Evaluation of a DNA probe of plasmid origin for the detection of Chlamydia trachomatis in cultures and clinical specimens.
This study evaluates five cryptic plasmid-derived DNA probes in a 4-h slot-blot hybridization assay for the detection of Chlamydia trachomatis in cultures and clinical specimens. The probes, consisting of either the entire cloned 7.5 kbp cryptic plasmid pSE8 or one of four Hin dIII/Eco RI fragments measuring 710, 1055, 710, and 500 bp, respectively, were labelled with Photoprobe biotin. The probe was detected using a streptavidin-alkaline phosphatase conjugate followed by addition of BCIP and NBT. The sensitivity of the assay, using 25 ng of probe DNA, ranged from 50 pg (with the entire plasmid as probe) to 5 ng (with the 500 bp fragment as probe). A total of 103 reference strains of Chlamydia trachomatis and other bacteria were tested for reactivity with the probes. All 18 reference strains of C. trachomatis hybridized with the probes. None of the DNA from the heterologous organisms tested was found to hybridize with any of the probes. A total of 174 samples taken from patients visiting the STD clinic at the University Hospital, University of Seville were used in the study. The overall sensitivity of the assay, using the 710 bp biotinylated probe was 94.5% compared to culture while the specificity was 97.5%. Positive and negative predictive values were 96.5% and 97.5%, respectively. It appears that the plasmid-derived probes used in this study could serve as useful tools for the rapid and specific detection of Chlamydia trachomatis in cell cultures and clinical specimens.
Cano
R J
RJ
Biological Sciences Department, California Polytechnic University, San Luis Obispo 93407.
Murrieta
C M
CM
Spaulding
D C
DC
Pascual
A
A
D001419:Bacteria; D015139:Blotting, Southern; D002478:Cells, Cultured; D002690:Chlamydia Infections / Q000175:diagnosis*; D002692:Chlamydia trachomatis / Q000302:isolation & purification*; D003001:Cloning, Molecular; D015342:DNA Probes; D004269:DNA, Bacterial; D005069:Evaluation Studies as Topic; D005260:Female; D015723:Gene Library; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics*; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D004269:DNA, Bacterial