Development of a Polymerase Chain Reaction Assay for the Detection of Haemophilus ducreyi

Johnson, Steven Ross; Martín, David H.; Cammarata, Catherine L.; Morse, Stephen

doi:10.1097/00007435-199401000-00004

Cited by 44 publications

(22 citation statements)

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“…Primer sets targeting the 16S rRNA gene (174)(175)(176), the ribosomal intergenic spacer region (177), recD (178,179), and the heat shock protein gene groEL (179,180) have been published.…”

Section: Identification By Pcrmentioning

confidence: 99%

Classification, Identification, and Clinical Significance of Haemophilus and Aggregatibacter Species with Host Specificity for Humans

2014

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SUMMARY The aim of this review is to provide a comprehensive update on the current classification and identification of Haemophilus and Aggregatibacter species with exclusive or predominant host specificity for humans. Haemophilus influenzae and some of the other Haemophilus species are commonly encountered in the clinical microbiology laboratory and demonstrate a wide range of pathogenicity, from life-threatening invasive disease to respiratory infections to a nonpathogenic, commensal lifestyle. New species of Haemophilus have been described ( Haemophilus pittmaniae and Haemophilus sputorum ), and the new genus Aggregatibacter was created to accommodate some former Haemophilus and Actinobacillus species ( Aggregatibacter aphrophilus , Aggregatibacter segnis , and Aggregatibacter actinomycetemcomitans ). Aggregatibacter species are now a dominant etiology of infective endocarditis caused by fastidious organisms (HACEK endocarditis), and A. aphrophilus has emerged as an important cause of brain abscesses. Correct identification of Haemophilus and Aggregatibacter species based on phenotypic characterization can be challenging. It has become clear that 15 to 20% of presumptive H. influenzae isolates from the respiratory tracts of healthy individuals do not belong to this species but represent nonhemolytic variants of Haemophilus haemolyticus . Due to the limited pathogenicity of H. haemolyticus , the proportion of misidentified strains may be lower in clinical samples, but even among invasive strains, a misidentification rate of 0.5 to 2% can be found. Several methods have been investigated for differentiation of H. influenzae from its less pathogenic relatives, but a simple method for reliable discrimination is not available. With the implementation of identification by matrix-assisted laser desorption ionization–time of flight mass spectrometry, the more rarely encountered species of Haemophilus and Aggregatibacter will increasingly be identified in clinical microbiology practice. However, identification of some strains will still be problematic, necessitating DNA sequencing of multiple housekeeping gene fragments or full-length 16S rRNA genes.

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“…Primer sets targeting the 16S rRNA gene (174)(175)(176), the ribosomal intergenic spacer region (177), recD (178,179), and the heat shock protein gene groEL (179,180) have been published.…”

Section: Identification By Pcrmentioning

confidence: 99%

Classification, Identification, and Clinical Significance of Haemophilus and Aggregatibacter Species with Host Specificity for Humans

2014

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“…The methods used to estimate the sensitivity and speci® city of these techniques are also different. DNA sequences of an anonymous cloned fragment of H. ducreyi was used as a target for a PCR assay 16 . An ampli® ed fragment of 1.1 kb was detected by Southern blotting and hybridization to a 32 P labelled 1.1 kb probe 16 .…”

Section: Discussionmentioning

confidence: 99%

Development of a heminested polymerase chain reaction assay for the detection of Haemophilus ducreyi in clinical specimens

et al. 2001

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Detection of Haemophilus ducreyi in genital ulcer specimens by culture lacks sensitivity. To enhance detection, a heminested polymerase chain reaction (PCR) assay was developed targeting the nucleotide sequence of a gene, designated p27, which encodes for a 27 kDa H. ducreyi-specific protein. The p27 PCR assay detected all (37/37) H. ducreyi strains tested and gave no amplified product from DNA extracts of any of 31 other microorganisms, from 30 non-genital ulcer specimens, or from 29 urethral and vaginal swab specimens collected from non-chancroid STD patients. In genital ulcer disease specimens, compared to combined positive results obtained by culture and a previously described PCR assay, the p27 PCR assay showed a sensitivity of 91% (48/53). The p27 PCR assay provides a specific and a sensitive detection of H. ducreyi in clinical specimens.

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“…A 1100 bp PCR product was detected by Southern blotting and hybridization to a 32P-labelled probe consisting of the entire sequence. In a first study (Johnson et al, 1994), they observed a sensitivity of 62% and a specificity of 52% relative to culture techniques. By modifying sample preparation, they found a sensitivity of 100% and a specificity of 84% compared with culturing (Johnson et al, 1995).…”

Section: Detection Of H Ducreyi By Pcrmentioning

confidence: 97%

“…Chui Depending on the number of cycles of amplification, they obtained sensitivities of 83% and 98% and specificities of 67% and 51% relative to culture. Johnson et al (1994Johnson et al ( , 1995 developed a PCR assay with two primers selected from the sequences of an anonymous fragment of DNA cloned from H . ducreyi.…”

Section: Detection Of H Ducreyi By Pcrmentioning

confidence: 99%

The rrs (16S)-rrl (23S) ribosomal intergenic spacer region as a target for the detection of Haemophilus ducreyi by a heminested-PCR assay

Rossau

Jannes

et al. 1998

Microbiology

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The intergenic spacer region between the rrs and rrl ribosomal RNA genes of Haemophilus ducreyi was analysed and the DNA sequence was used for the selection of specific PCR primers. A highly sensitive and specific heminested-PCR assay for the identification of H. ducreyi was developed. The assay showed a sensitivity of 96% on genital ulcer specimens from patients with clinically diagnosed chancroid, compared with a sensitivity of 56% for culture methods.These results indicate that this PCR assay has the potential to become an accurate and easy reference method for the detection of H. ducreyi.

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Development of a Polymerase Chain Reaction Assay for the Detection of Haemophilus ducreyi

Cited by 44 publications

References 0 publications

Classification, Identification, and Clinical Significance of Haemophilus and Aggregatibacter Species with Host Specificity for Humans

Classification, Identification, and Clinical Significance of Haemophilus and Aggregatibacter Species with Host Specificity for Humans

Development of a heminested polymerase chain reaction assay for the detection of Haemophilus ducreyi in clinical specimens

The rrs (16S)-rrl (23S) ribosomal intergenic spacer region as a target for the detection of Haemophilus ducreyi by a heminested-PCR assay

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