Establishment of a particle-counting method for purified elementary bodies of chlamydiae and evaluation of sensitivities of the IDEIA Chlamydia kit and DNA probe by using the purified elementary bodies.
To evaluate the sensitivity of commercially available test kits for detection of chlamydiae, we established a method of purifying Chlamydia trachomatis and Chlamydia pneumoniae elementary bodies (EBs). We then subjected the purified EBs, together with the purified EBs of Chlamydia psittaci, to the IDEIA Chlamydia (IDEIA) and DNA probe test kits to determine the EB numbers at the detection limits. The sensitivities of the test kits were thus compared. The results can be summarized as follows. (i) Intact EBs in the purified preparations were present at 100, 96.3, and 97% for the C. psittaci Cal 10, C. trachomatis L2/434/Bu (L2), and C. pneumoniae TW-183 strains, respectively. The preparations of the L2 and TW-183 EBs contained a few EB envelopes, which reacted with antilipopolysaccharide monoclonal antibodies, as did the intact EBs, indicating that elimination of EB envelopes is not required for testing of the IDEIA kit's sensitivity. (ii) We established a method of counting intact EBs and EB envelopes under a scanning electron microscope after sedimentation of EBs on a coverslip by centrifugation. (iii) The EB numbers per assay at the cutoff level, which is set up in the IDEIA kit, were 9.6 x 10(2), 6.5 x 10(3), and 2.5 x 10(4) for the L2, TW-183, and Cal 10 strains, respectively. When the same EB preparations were applied to the DNA probe kit, the EB number at the cutoff level was 7.5 x 10(3) per assay for the L2 strain, but no reaction occurred for the Cal 10 and TW-183 strains at any EB number, indicating that the DNA probe kit is highly specific for C. trachomatis. Although the IDEIA kit designed for detection of C. trachomatis showed a sensitivity superior to that of the DNA probe, the chlamydial species was not determined by the IDEIA kit.
Miyashita
N
N
Department of Microbiology, Kawasaki Medical School, Okayama, Japan.
Matsumoto
A
A
D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D002689:Chlamydia / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002690:Chlamydia Infections / Q000175:diagnosis; D002692:Chlamydia trachomatis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D015342:DNA Probes; D005069:Evaluation Studies as Topic; D006801:Humans; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D013045:Species Specificity
D015342:DNA Probes