Detection of<i>Chlamydia trachomatis</i>by nucleic acid sandwich hybridization

Palva, Airi; Jousimies‐Somer, Hannele; Saikku, Pekka; Väänänen, Pertti; Söderlund, Hans; Ranki, Marjut

doi:10.1111/j.1574-6968.1984.tb01040.x

Cited by 54 publications

(5 citation statements)

References 32 publications

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“…These sensitivity levels are similar to those which have been reported for other DNA probes using radioisotopic detection methods [8,9] and are comparable with levels obtainable with fluorescent antibody staining [11]. However, in cases where the infecting species is unknown (suspected C. pneumoniae infection) the genus specific probe, pFEN207 should be used.…”

Section: Psittaci Infections In Birds or In Humans Withsupporting

confidence: 81%

Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Rasmussen

1991

FEMS Microbiology Letters

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Fewer than 10(5) elementary bodies of Chlamydia psittaci could be detected by using DNA hybridisation with a plasmid probe specific for avian chlamydial strains. PCR amplification of chlamydial DNA using primers specific for conserved regions of the major outer membrane protein gene enabled the detection of fewer than 10 elementary bodies. DNA could be amplified from 22 of the 24 chlamydial strains tested including avian, feline, ovine, caprine, koala and lymphogranuloma venereum strains.

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Section: Psittaci Infections In Birds or In Humans Withsupporting

confidence: 81%

Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Rasmussen

1991

FEMS Microbiology Letters

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“…After the sensitivity and specificity of the method proposed for detection of chlamydia in 1984 [16] had been markedly improved, it is widely used in the diagnostics of chlamydial infections [8,11,19]. …”

Section: Abstract: Biotin-labeled Dna Probe; Dot Hybridization; C Tmentioning

confidence: 99%

Diagnosis of urogenital chlamydiasis by dot hybridization with biotin-labeled DNA probe

et al. 1996

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The DNA fragment of C. trachomatis cryptic plasmid (1115 bp long) is cloned into pBR322vector. The recombinant plasmid pCTpl binds to purified chlamydial DNA and DNA isolated from epithelial cells of patients with clinical manifestations of chlamydiasis and does did not bind to DNA of M. hominis, U. urealyticum, E. coli, herpes simplex virus, and human DNA. The probe sensitivity is 64.5%, its specificity is 84.9%, and the specificities for positive and negative results are 79.2 and 85.7%, respectively, compared with the polymerase chain reaction. These parameters are 61,6, 81.4, 78.2, and 85.6%, respectively, compared with the immunofluorescent method. Key Words: biotin-labeled DNA probe; dot hybridization; C. trachomatis; diagnosis of chlamydiasisChlamydia trachomatis is a widespread sexually-transmitted pathogenic microorganism. Chlamydial infection may take an asymptomatic, chronic, or acute course with numerous clinical manifestations involving the reproductive system, eyes, kidneys, liver, lymph vessels, and joints [1,2,9,10,15]. Chlamydia can persist for many years in the body causing no disease and exhibiting no antigenic activity. These specific features of chlamydial infection hamper its clinical and serologic diagnosis.Diagnostic methods based on specificity of genetic material of a causative agent (including DNA hybridization) have found wide application in recent years. After the sensitivity and specificity of the method proposed for detection of chlamydia in 1984 [16] had been markedly improved, it is widely used in the diagnostics of chlamydial infections [8,11,19]. MATERIALS AND METHODSEpithelial cells from the urethra (males) and cervical canal (females) were used in the study. Cells were collected with a Medscand or Abbott devices. Processing of clinical material, DNA isolation, and dot hybridization were carried out as described previously [3]. The ceils were washed with normal saline and lysed with 1% SDS and 100 gg/ml proteinase K. DNA was precipitated with 2 volumes of 96% ethanol, dissolved in 10 volumes of SSC buffer (1.5 M NaC1 and 0.15 M sodium citrate), degraded by boiling, and transferred onto a nitrocellulose filter. Hybridization with biotin-labeled probe was carried out in a 50% aqueous solution of formamide for 14-16 h at

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“…The sandwich hybridization technique has been described by Ranki et al [9] and has later been developed into a diagnostic technique by Palva et al [10]. The method does not require the DNA to be radiolabelled, and no purification of the material is necessary.…”

Section: Introductionmentioning

confidence: 99%

Exceptionally high copy numbers of a staphylococcal plasmid in Bacillus subtilis revealed by a sandwich hybridization technique

Nyberg¹

1985

FEMS Microbiology Letters

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The copy number of a pUB110 derivative, pKTH10, containing the α‐amylase gene from Bacillus amyloliquefaciens, was determined, using an assay based on a sandwich hybridization technique. In this method, a known gene on the plasmid is hybridized between two non‐overlapping fragments of that same gene, cloned into separate vectors. One fragment is used as a radiolabelled probe and the other bound to a filter, forming a three‐component, ‘sandwich’ hybrid when the relevant gene is present in the sample. Since the hybridization can only take place in the presence of the relevant gene, the amount of radioactivity binding to the filters will be proportional to the concentration of this gene in the sample. We utilized the α‐amylase gene on the plasmid to form the sandwich hybrid. The copy number was of a totally different magnitude from what has previously been reported, and ranged from 2500 copies/viable cell in early logrithimic growth phase to about 500 in late stationary phase.

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Detection ofChlamydia trachomatisby nucleic acid sandwich hybridization

Cited by 54 publications

References 32 publications

Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Diagnosis of urogenital chlamydiasis by dot hybridization with biotin-labeled DNA probe

Exceptionally high copy numbers of a staphylococcal plasmid in Bacillus subtilis revealed by a sandwich hybridization technique

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