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Evaluation of a fluorescent DNA hybridization assay for the detection ofNeisseria gonorrhoeae

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Abstract

This study evaluates a four-hour fluorescent DNA hybridization assay using both known bacterial isolates and clinical specimens. A biotinylated oligonucleotide probe from a sequence of the plasmid-encoded genecppB was used. Hybrids were detected by addition of a streptavidin-alkaline phosphatase conjugate, followed by incubation for 30 min in a fluorescent substrate for alkaline phosphatase. The level of detection of the fluorescent assay was 0.1 pg of cryptic plasmid DNA or 200 cfu of the plasmid-containing strain NG 34/85 ofNeisseria gonorrhoeae. A total of 119 reference strains ofNeisseria gonorrhoeae and other related bacteria were tested for reactivity with the probe. AllNeisseria gonorrhoeae strains, including eight plasmid-free strains, hybridized with the probe. Fluorescence ratios were 2.67 for plasmid-free strains and 3.85 for plasmid-containing strains. Of the heterologous microorganisms tested, only one of six strains ofNeisseria cinerea gave a fluorescence ratio above the 2.0 cut-off value for positivity with the probe at a cell density of 1 × 104 cfu. The probe was also evaluated using clinical specimens from 100 patients attending a clinic for sexually transmitted diseases. The sensitivity of the assay was 100 % while the specificity was 97.5 %. Positive and negative predictive values were 91.2 % and 100 %, respectively. The fluorescent DNA hybridization assay for the detection ofNeisseria gonorrhoeae described here thus appears to be a highly specific and sensitive assay.

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Authors and Affiliations

  1. Biological Sciences Department, California Polytechnic State University, 93407, San Luis Obispo, California, USA

    R. J. Cano

  2. Department of Microbiology, University Hospital, Seville, Spain

    J. C. Palomares & M. J. Torres

  3. JBL Scientific, 277 Granada Drive, 93401, San Luis Obispo, California, USA

    R. E. Klem

Authors
  1. R. J. Cano
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  2. J. C. Palomares
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  3. M. J. Torres
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  4. R. E. Klem
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Cano, R.J., Palomares, J.C., Torres, M.J. et al. Evaluation of a fluorescent DNA hybridization assay for the detection ofNeisseria gonorrhoeae . Eur. J. Clin. Microbiol. Infect. Dis. 11, 602–609 (1992). https://doi.org/10.1007/BF01961666

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