Construction of DNA Probes for the Identification of Haemophilus ducreyi

Parsons, Linda M.; Shayegani, Mehdi; Waring, Alfred L.; Bopp, Lawrence H.

doi:10.1016/b978-0-12-463000-0.50009-8

Cited by 9 publications

(9 citation statements)

References 57 publications

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“…These signature nucleotides have been exploited in nucleic-acid-based identification systems, directed against specific rRNA sequences, which are species-or subspecies-specific. Nucleic-acid-based detection systems have been developed for Mycoplasma (Gobel et al, 1987), Treponema (Jensen et al, 1990), Campylobacter (Cox et al, 1990), Haemophilus (Parsons et al, 1989), Borrelia and Clostridium (Wilson et al, 1988).…”

Section: Resultsmentioning

confidence: 99%

Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination

Marconi¹,

Garon²

1992

Journal of General Microbiology

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As part of a continuing effort to assess genetic variation among isolates of Borrelia burghrferi we have determined the 16s rRNA signature nucleotide makeup of two tick isolates from the USSR. Signature nucleotides were identified via reverse transcriptase primer extension sequencing of select regions of the 16s rRNA molecule. In addition, the near complete 16s rRNA sequence of one of the isolates, R-IP3, was determined and utilized in a phylogenetic assessment. The sequence was aligned with the 16s rRNA sequences of other B. burghrferi isolates as well as with other B o r r e l . species. Distance matrix analyses were performed and a phylogenetic tree was constructed. These analyses demonstrate that these isolates belong to a third previously unidentified genomic group of B. burghrferi.

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Section: Resultsmentioning

confidence: 99%

Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination

Marconi¹,

Garon²

1992

Journal of General Microbiology

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“…With the introduction of DNA diagnostic methods (17) and the ability to amplify the signal with the polymerase chain reaction (PCR), newer tests should improve the sensitivity and specificity of diagnostic tests for chancroid. First-generation DNA probes for H. ducreyi have been reported with sensitivities of 103 to 104 organisms and 100% specificity (25,26). We report here our results with the development of primer sets and probes for the diagnosis of chancroid by DNA amplification using PCR.…”

mentioning

confidence: 75%

Development of the polymerase chain reaction for diagnosis of chancroid

et al. 1993

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The published nucleotide sequences of the 16S rRNA gene of Haemophilus ducreyi were used to develop primer sets and probes for the diagnosis of chancroid by polymerase chain reaction (PCR) DNA amplification. One set of broad specificity primers yielded a 303-bp PCR product from all bacteria tested. Two 16-base probes internal to this sequence were species specific for H. ducreyi when tested with 12 species of the families PasteureUaceae and Enterobacteriaceae. The two probes in combination with the broad specificity primers were 100% sensitive with 51 strains of H. ducreyi isolated from six continents over a 15-year period. The direct detection of H. ducreyi from 100 clinical specimens by PCR showed a sensitivity of 83 to 98% and a specificity of 51 to 67%, depending on the number of amplification cycles.

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“…Parsons et al evaluated the ability of three 32 P labelled DNA probes to hybridise with H ducreyi DNA in both bacterial suspensions and infected rabbit lesion material blotted onto nitrocellulose membranes. 44 The probes reliably detected 10 4 CFU of H ducreyi in pure and mixed cultures. The sensitivity of this technique for diagnosis of chancroid would be greatly increased if initial amplification of H ducreyi DNA in the specimen could be made possible either by direct bacterial growth or by a DNA amplification based methodology.…”

Section: Nucleic Acid Probe Technologymentioning

confidence: 95%

Diagnostic tests for chancroid

Lewis

2000

Sexually Transmitted Infections

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Construction of DNA Probes for the Identification of Haemophilus ducreyi

Cited by 9 publications

References 57 publications

Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination

Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination

Development of the polymerase chain reaction for diagnosis of chancroid

Diagnostic tests for chancroid

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