Evaluation of a prototype DNA probe test for the noncultural diagnosis of gonorrhea

Granato, Paul A.; Franz, M.

doi:10.1128/jcm.27.4.632-635.1989

Cited by 36 publications

(6 citation statements)

References 20 publications

(26 reference statements)

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“…In contrast to these limitations, the nonisotopic probe is safe and has a considerably longer shelf life; the AE-DNA probe can be stored for over 1 year if it is refrigerated. An acridinium ester has already been used to label the DNA probe for Neisseria gonorrhoeae (the prototype Gen-Probe PACE system) (5). However, in that assay, magnetic particles that specifically bind to hybridized probes are used, and washing must be repeated three times to remove unhybridized probes.…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

et al. 1991

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The detectability of mycobacteria in culture by the use of nonisotopic, chemiluminescent DNA probes for Mycobacterium tuberculosis and the M. avium-M. intracellulare complex (MAC) was evaluated and compared with that by the use of 125I-labeled DNA probes for the same mycobacteria. In the assay, rRNA-directed DNA probes labeled with acridinium ester (AE-DNA probes) were used. Unhybridized probes were chemically degraded, and the esterified acridinium on the hybridized probes was hydrolyzed by the addition of alkaline hydrogen peroxide solution, resulting in the production of visible light which was measured with a luminometer. The detection limits of the AE-DNA probes were almost the same as those of the 1251-labeled DNA probes. A total of 107 clinical isolates of mycobacteria (47 isolates of M. tuberculosis, 36 MAC, and 24 atypical mycobacteria other than MAC) were tested. The sensitivity and specificity of the AE-DNA probes for M. tuberculosis were 100% both for the conventional method and with the 125I-labeled DNA probe. The sensitivity and specificity of the AE-DNA probes for MAC were 97.2 and 100%, respectively, for the conventional method and were both 100% with the 125I-labeled DNA probes. Because the procedure is simple, reliable, rapid (it can be completed within an hour), and safe (it does not use radioisotopes), it can easily be performed in any clinical laboratory.

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Section: Discussionmentioning

confidence: 99%

“…In some of them, such as those for Legionella spp. (4), N. gonorrhoeae (5), and Chlamydia trachomatis (8), organisms can be detected directly in clinical specimens by the DNA probes. The detection limits of the AE-DNA probes for mycobacteria achieved in this study were at the level of 106 CFU.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

et al. 1991

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“…Conventional culture procedures are dependent on the presence of viable bacteria for microbial isolation. Nonculture alternatives for the laboratory detection of Neisseria gonorrhoeae in urogenital specimens include enzyme immunoassay (11) and nucleic acid hybridization (4,12).…”

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confidence: 99%

Comparison of Gen-Probe DNA probe test and culture for the detection of Neisseria gonorrhoeae in endocervical specimens

et al. 1991

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A 2-h nonisotopic DNA probe assay for the direct detection of Neisseria gonorrhoeae in urogenital specimens has recently been modified (PACE 2; Gen-Probe, San Diego, Calif.). The new assay format was developed to increase the sensitivity of the assay and simplify procedural steps. In this study, the new DNA probe test was compared with a culture reference method for the detection of N. gonorrhoeae in endocervical specimens. The results of the DNA probe test were expressed as a ratio of relative light units (RLU) of the specimen/RLU of the cutoff recommended by the manufacturer. All patient samples with sample RLU/cutoff RLU ratios less than 0.7 were interpreted as negative, and ratios greater than 2.0 were interpreted as positive for gonorrhea. Samples with sample RLU/cutoff RLU ratios between 0.7 and 2.0 were repeated until two or more consistent negative or positive ratios were obtained. A total of 469 specimens were tested with an overall disease prevalence of 6.1%. Of the 469 patients tested, 5 specimens (1.0%) fell in this borderline region and were retested. If the manufacturer's recommended cutoff value had been used, the original DNA probe results would have resulted in two false-positives. Our data were analyzed for both symptomatic (prevalence, 11.7%) and asymptomatic (prevalence, 2%) women. The study indicated that with our modification of the manufacturer's endpoint interpretation, the DNA probe test was essentially equivalent to the culture method in terms of sensitivity, specificity, and positive and negative predictive values in both symptomatic and asymptomatic patient populations. The new DNA probe test can serve as a suitable screening and diagnostic test for the diagnosis of gonorrheal genital infections in women. Additionally, it offers the advantages of rapid turnaround time and ease of use and allows simultaneous testing for Chlamydia trachomatis on the same specimen. Gonorrhea remains one of the most important sexually transmitted diseases worldwide, with a reported 720,000 cases in the United States in 1988 (1). Gynecologic infections can range from asymptomatic to mildly symptomatic to severe, with long-term serious sequelae including infertility, chronic pelvic pain, and ectopic pregnancy. It has also been implicated in obstetric complications, neonatal pneumonia, and conjunctivitis. The control of epidemic infections and their sequelae remains dependent on correct and timely diagnosis.

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“…Noncultural techniques for diagnosing sexually transmitted diseases and, in particular, chlamydial and gonorrheal infections are now widely used. Such techniques are largely based on either immunofluorescence microscopy, enzyme immunoassay (1,3,12,14), or nucleic acid hybridization (4,10,11,15). These nonculture methods offer several advantages over conventional culture procedures in that the diagnosis is not dependent on the presence of viable microorganisms for microbial isolation and that turnaround times can be significantly reduced.…”

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confidence: 99%

Value of a DNA probe assay (Gen-Probe) compared with that of culture for diagnosis of gonococcal infection

et al. 1993

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The Gen-Probe PACE 2 system for Neisseria gonorrhoeae (GP), which uses a chemiluminescently labeled DNA probe, was compared with conventional culture as the method of reference. A total of 1,750 specimens were collected from 496 females and 623 males visiting the outpatient clinic of the Sexually Transmitted Diseases Department of the Westeinde Hospital, The Hague, The Netherlands, during the year 1991. The prevalences of gonorrhea culture-positive men and women were 14.9 and 7.7%o, respectively. The overall positive rate was 8.7%. Sensitivity, specificity, and positive and negative predictive values of GP were 97.1, 99.1, 90.6, and 99.8%, respectively. A total of 12 of 13 patients with positive GP results and negative cultures may have had a gonococcal infection, a conclusion based on clinical symptoms, positive methylene blue smears, and high relative light unit ratios. The DNA probe test can be useful as a suitable screening and diagnostic test for gonorrheal infection in men and women. An advantage of using this DNA probe technique is that simultaneous testing for Chlamydia trachomatis of the same specimen is possible. We also examined whether (all) rRNA had disappeared after adequate treatment for gonococcal and/or chlamydial infection in 30 patients. None of those positive patients showed a positive result in the DNA probe assay after treatment.

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Evaluation of a prototype DNA probe test for the noncultural diagnosis of gonorrhea

Cited by 36 publications

References 20 publications

Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

Comparison of Gen-Probe DNA probe test and culture for the detection of Neisseria gonorrhoeae in endocervical specimens

Value of a DNA probe assay (Gen-Probe) compared with that of culture for diagnosis of gonococcal infection

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