Background:
@En Chlamydia trachomatis is an important cause of sexually transmitted disease. Diagnostic tests based on the recognition of DNA sequences have been developed. The DNA probe assay has been described to detect specific parts of chlamydial nucleic
acids.
We evaluated the gene probe(pFEN50) encoding the MOMP of C.trachomatis for detection of trachomatis from cervical specimens of 80 patients with pelvic inflammatory disease.
@ES Methods
@EN Chlamydial culture was performed on cyclohexamide(2㎍/ml)-treated McCoy cell monolayer in shell vials, and the specimens were spotted onto Nytran-plus after boiling in lysing buffer(50mM Tris-HCI, pH 7.5; 1% Triton X-100; 1mM EDTA, Proteinase
K
400
g/ml). Hybridization was then carried out.
@ES Results:
@EN The nucleic acid(pFEN50) was hybridized with one 9.8 kb fragment from Bam HI-cleaved C.trachomatis(L2) DNA. The sensitivity of the pFEN50 was 100 pg DNA. No binding of the pFEN50 to C. pneumoniae(TWAR) and other microorganisms(Staphylococcus
aureus,
coagulase negative staphylococcus, Grams-positive bacilli, α-hemolytic streptococcus, β-hemolytic streptococcus, Enterobacter cloacae, Acinetobacter lwoffi, Pseudomonas aeruginosae, Escherichia coli, Klebsiella pneumoniae) was observed. Tree
were
positive for chlamydial culture and dot-blot hybridization. Four of eighty cases showed inconsistent results. Three of the four cases were culture negative, gene probe positive, which might be explained as false negativity of culture. One case
was
culture positive and gene probe negative, which might be due to the presence of dot-blot hybridization inhibitors in sample or the absence of inclusion bodies.
@ES Conclusion
@EN We conclude that the dot-blot hybridization assay can be used to demonstrate the presence of C.trachomatis in many clinical specimens simultaneously.
In addition, the pFEN50 nucleic acid is a highly sensitive and specific gene probe for the detection of all serovar of C. trachomatis in clinical specimens.
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