Development of urease conjugated enzyme-linked immunosorbent assays (ELISA) for the detection of IgM and IgG antibodies against Mycoplasma pneumoniae in human sera

Chia, W. K.; Spence, L.; Dunkley, Lisa; Bradbury, W C

doi:10.1016/0732-8893(88)90078-8

Cited by 11 publications

(5 citation statements)

References 17 publications

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“…The final supernatant was precipitated with a final concentration of 3 M sodium acetate (NaOAC) pH 7.5 and 2 volumes ethanol added. The samples were placed at -20°C for 12 h. DNA was recovered following a 10 min centrifugation at 12000 g, 4"C, followed by a wash in 70% ethanol, centrifugation as before and resuspension of the DNA pellet in 100 p1 Tris/EDTA (10 Table I). Mp-1 was included for three-primer DNA amplification and confirmation that the amplified products were indeed M .…”

Section: N a E X T R A C T I O Nmentioning

confidence: 99%

The detection of Mycoplasma pneumoniae in nasal polyps

et al. 1996

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The aetiology and microbial flora of nasal polyps is not well understood. No study in the literature has reported an association between the sub-bacterium Mycoplasma pneumoniae and nasal polyps. We have developed an assay method using the Polymerase Chain Reaction (PCR) to amplify a specific region of the M. pneumoniae DNA in extracts of clinical samples using species-specific primers designed to a region of the 16S rRNA. The presence of M. pneumoniae was detected in 13/14 (93%) nasal polyps, in 4/5 (80%) rhinosinusitis mucosal samples but only in 1/7 (14%) of control samples (obstructive turbinates). An epidemic of infections due to M. pneumoniae is expected to occur in 1995. We believe this assay could form the basis of a rapid technique for M. pneumoniae detection. We also propose that the presence of M. pneumoniae may be of importance in the aetiology of nasal polyps.

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Section: N a E X T R A C T I O Nmentioning

confidence: 99%

The detection of Mycoplasma pneumoniae in nasal polyps

et al. 1996

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“…Culture is time-consuming, taking several weeks to produce results and is relatively insensitive (Dorigo-Zetsma et al, 1999;Harris et al, 1988;Kok et al, 1988). Serological methods are insufficiently sensitive and require paired serum samples from the acute and convalescent phases of the disease, thus only allowing a retrospective diagnosis (Dorigo-Zetsma et al, 1999;Chia et al, 1988;Sillis, 1990;Fedorko et al, 1995). More rapid, higher sensitivity methods were therefore developed, one of them being the PCR for fragments of the P1 gene or the 16S rRNA gene (Dorigo-Zetsma et al, 1999;Tjhie et al, 1994;Ieven et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae

Saito

Misawa

Moriya

et al. 2005

Journal of Medical Microbiology

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A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Mycoplasma pneumoniae was developed and evaluated. The assay specifically amplified only M. pneumoniae sequences, and no cross-reactivity was observed for other Mycoplasma species or respiratory bacterial species. The detection limit for this assay was found to be 2 3 10 2 copies, corresponding to 2-20 colour changing units of M. pneumoniae in 1 h, as observed in a real-time turbidimeter and electrophoretic analysis. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis as well as direct sequencing of the amplified product. The assay was applied to 95 nasopharyngeal swab samples collected from patients or from healthy individuals, and compared to a real-time PCR assay in-house. A concordance of 100 % was observed between the two assays. The LAMP assay is easy to perform, shows a rapid reaction and is inexpensive. It may therefore be applied in the routine diagnosis of M. pneumoniae infection in the clinical laboratory.

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“…An accurate diagnosis with convalescentphase samples is often made many days after the onset of disease (28). Sensitivity and specificity values are between 55 and 100%, depending on the serological method used and the patient population tested (1,4,8,10,15,18,25). PCR has been shown to offer the potential of increased sensitivity and rapidity compared to other diagnostic tests.…”

mentioning

confidence: 99%

Comparison and Evaluation of Real-Time PCR, Real-Time Nucleic Acid Sequence-Based Amplification, Conventional PCR, and Serology for Diagnosis of Mycoplasma pneumoniae

Templeton¹,

Scheltinga²,

Graffelman³

et al. 2003

J Clin Microbiol

134

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Mycoplasma pneumoniae is a common cause of community-acquired pneumonia and lower-respiratory-tract infections. Diagnosis has traditionally been obtained by serological diagnosis, but increasingly, molecular techniques have been applied. However, the number of studies actually comparing these assays is limited. The development of a novel duplex real-time PCR assay for detection of M. pneumoniae in the presence of an internal control real-time PCR is described. In addition, real-time nucleic acid sequence-based amplification (NASBA) on an iCycler apparatus is evaluated. These assays were compared to serology and a conventional PCR assay for 106 clinical samples from patients with lower-respiratory-tract infection. Of the 106 samples, 12 (11.3%) were positive by all the molecular methods whereas serology with acute sample and convalescent samples detected 6 (5.6%) and 9 (8.5%), respectively. Clinical symptoms of the patients with Mycoplasmapositive results were compared to those of the other patients with lower-respiratory-tract infections, and it was found that the results for mean lower age numbers as well as the presence of chills, increased erythrocyte sedimentation rate, and raised C-reactive protein levels showed significant differences. Molecular methods are superior for diagnosis of M. pneumoniae, providing more timely diagnosis. In addition, using real-time methods involves less hands-on time and affords the ability to monitor the reaction in the same tube.

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Development of urease conjugated enzyme-linked immunosorbent assays (ELISA) for the detection of IgM and IgG antibodies against Mycoplasma pneumoniae in human sera

Cited by 11 publications

References 17 publications

The detection of Mycoplasma pneumoniae in nasal polyps

The detection of Mycoplasma pneumoniae in nasal polyps

Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae

Comparison and Evaluation of Real-Time PCR, Real-Time Nucleic Acid Sequence-Based Amplification, Conventional PCR, and Serology for Diagnosis of Mycoplasma pneumoniae

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