The specificity of pilin DNA sequences for the detection of pathogenic Neisseria

Kolberg, Janice A.; Besemer, Diana J.; Stempien, Michelle M.; Urdea, Mickey S.

doi:10.1016/0890-8508(89)90037-6

Cited by 7 publications

(3 citation statements)

References 26 publications

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“…Though this may have resulted in less inactivation of the alkaline phosphatase marker enzyme, they used the unamplified fluorogenic substrate 4-methylumbelliferone phosphate, which is less sensitive than the enzyme amplification detection method. The assay was also 10-to 1000-fold more sensitive than DNA-targeted bacterial detection systems using direct or indirect enzyme detection (21,22).…”

Section: Discussionmentioning

confidence: 99%

A Sandwich Hybridization Assay Employing Enzyme Amplification for termination of Specific Ribosomal RNA from Unpurified Cell Lysates

Wicks

Cook

Barer

et al. 1998

Analytical Biochemistry

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Section: Discussionmentioning

confidence: 99%

A Sandwich Hybridization Assay Employing Enzyme Amplification for termination of Specific Ribosomal RNA from Unpurified Cell Lysates

Wicks

Cook

Barer

et al. 1998

Analytical Biochemistry

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“…It has been shown in several studies that plasmidfree strains of Neisseria gonorrhoeae may possess cryptic plasmid sequences integrated in the bacterial chromosome (15,16,19). This observation may explain the reactivity of the probe with all the strains ofNeisseria gonorrhoeae tested, regardless of the presence of the cryptic plasmid in the strains tested.…”

Section: Discussionmentioning

confidence: 76%

“…The detection of this organism in the clinical laboratory is still largely dependent upon traditional methods (2), although a number of alternative methods have been developed for the rapid and specific detection of Neisseria gonorrhoeae in clinical specimens. These include antigen detection techniques (3-5) and DNA hybridization assays (6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) in clinical specimens. When used as culture confirmation tests the sensitivity and specificity of these assays were in the range of 91-100 % (6-8, 15, 16), When used for direct detection of gonococci in clinical specimens, however, these values were in the range of 81-95 % (9-15).…”

mentioning

confidence: 99%

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

Cano

Palomares

Torres

et al. 1992

Eur. J. Clin. Microbiol. Infect. Dis.

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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 gene cppB 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 of Neisseria gonorrhoeae. A total of 119 reference strains of Neisseria gonorrhoeae and other related bacteria were tested for reactivity with the probe. All Neisseria 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 of Neisseria cinerea gave a fluorescence ratio above the 2.0 cut-off value for positivity with the probe at a cell density of 1 x 10(4) 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 of Neisseria gonorrhoeae described here thus appears to be a highly specific and sensitive assay.

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Controlled Synthetic Oligonucleotide Networks for the Detection of Pathogenic Organisms

Urdea¹

1991

Rapid Methods and Automation in Microbiology and Immunology

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The specificity of pilin DNA sequences for the detection of pathogenic Neisseria

Cited by 7 publications

References 26 publications

A Sandwich Hybridization Assay Employing Enzyme Amplification for termination of Specific Ribosomal RNA from Unpurified Cell Lysates

A Sandwich Hybridization Assay Employing Enzyme Amplification for termination of Specific Ribosomal RNA from Unpurified Cell Lysates

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

Controlled Synthetic Oligonucleotide Networks for the Detection of Pathogenic Organisms

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