Problems associated with serotyping strains of Ureaplasma urealyticum

Stemke, Gerald W.; Robertson, Janet A.

doi:10.1016/0732-8893(85)90005-7

Cited by 33 publications

(51 citation statements)

References 20 publications

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“…Although the results of gene-based methods are expected to be more easily reproducible than those of phenotyping, experience has taught us that typing cloned, laboratory-adapted strains is much easier than working with wildtype isolates (26). The serotyped isolates that were mba genotyped in this study emphasize this lesson.…”

Section: Discussionmentioning

confidence: 61%

Simple Method for Determining Biovar and Serovar Types of Ureaplasma urealyticum Clinical Isolates Using PCR–Single-Strand Conformation Polymorphism Analysis

Pitcher¹,

Sillis

Robertson

2001

J Clin Microbiol

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Ureaplasma urealyticum has been associated with urethritis in men, obstetric problems in women, and respiratory distress syndrome in preterm infants. U. urealyticum can be divided into two biovars comprising 14 serovars. Partial sequences of genes encoding the multiple-banded antigens of the cell surface are known. Using a commercially available precast DNA mutation detection gel system, we have developed a simple and reproducible PCR-single-strand conformation polymorphism analysis method for differentiating the biovars of this species that reveals five patterns among the 14 serovars and enables clinical isolates to be typed directly from broth cultures.

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Section: Discussionmentioning

confidence: 61%

Simple Method for Determining Biovar and Serovar Types of Ureaplasma urealyticum Clinical Isolates Using PCR–Single-Strand Conformation Polymorphism Analysis

Pitcher¹,

Sillis

Robertson

2001

J Clin Microbiol

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“…Similar to the species debate, there are numerous conflicting studies attempting to link a specific serovar with disease. This is further confounded through the use of flawed serotyping methodologies, many of which were reported shortly after the conception of the original serotyping scheme and showed multiple cross-reactions among individual serovars (9). Despite efforts to improve on this (10), disparities in serotyping methodologies have not been suitably resolved.…”

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confidence: 99%

High-Resolution Melt PCR Analysis for Genotyping of Ureaplasma parvum Isolates Directly from Clinical Samples

et al. 2014

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c Ureaplasma sp. infection in neonates and adults underlies a variety of disease pathologies. Of the two human Ureaplasma spp., Ureaplasma parvum is clinically the most common. We have developed a high-resolution melt (HRM) PCR assay for the differentiation of the four serovars of U. parvum in a single step. Currently U. parvum strains are separated into four serovars by sequencing the promoter and coding region of the multiple-banded antigen (MBA) gene. We designed primers to conserved sequences within this region for PCR amplification and HRM analysis to generate reproducible and distinct melt profiles that distinguish clonal representatives of serovars 1, 3, 6, and 14. Furthermore, our HRM PCR assay could classify DNA extracted from 74 known (MBA-sequenced) test strains with 100% accuracy. Importantly, HRM PCR was also able to identify U. parvum serovars directly from 16 clinical swabs. HRM PCR performed with DNA consisting of mixtures of combined known serovars yielded profiles that were easily distinguished from those for single-serovar controls. These profiles mirrored clinical samples that contained mixed serovars. Unfortunately, melt curve analysis software is not yet robust enough to identify the composition of mixed serovar samples, only that more than one serovar is present. HRM PCR provides a single-step, rapid, cost-effective means to differentiate the four serovars of U. parvum that did not amplify any of the known 10 serovars of Ureaplasma urealyticum tested in parallel. Choice of reaction reagents was found to be crucial to allow sufficient sensitivity to differentiate U. parvum serovars directly from clinical swabs rather than requiring cell enrichment using microbial culture techniques.

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“…The relationship between serovars and disease syndromes needs to be studied further (Grattard et al, 1995 ;Naessens et al, 1988). However, this has been limited by technical difficulties and cross-reactions associated with serotyping (Wiley & Quinn, 1984 ;Quinn et al, 1981 ;Robertson & Stemke, 1979 ;Stemke & Robertson, 1985), even when monoclonal antibodies were used (Naessens et al, 1998 CTG GTT TTG TTT CAA AAC CTA T427 UMA2A2* UU A\B 6 6 454CAC TTC CTG GTT TTG TTT CAA AAC431 UMA2A* UU A 66 479CCA CTT CCT GGT TTT GTA GTT TC457 UMA10A* UU B 68 479CCA CTT CCT GGT TGT GTA GTT GA457 UMA4A1* UU C 68 452TT GCC TGG TTG TGT TTC GAA CTC430 UMA4A2* UU C 68 454CAT TGC CTG GTT GTG TTT CGA AC432 UMA9A1* UU D 66 460CTG GAG TTG GTG TAG GCG CAT440 UMA9A2* UU D 66 462TTC TGG AGT TGG TGT AGG CGC442 UMA7A1 UU E 74 245GTA ATT GCA ACA TGG AAT TCA GTT TCA219 UMA7A2* UU E 66 458GGT TCT GGT GTA TGA GTG CTT TT436 UMA7A3* UU E 66 461GTT GGT TCT GGT GTA TGA GTG C440 * Primers designed specifically for this study. All others have been previously published (see text for references).…”

Section: Introductionmentioning

confidence: 99%

Molecular genotyping of human Ureaplasma species based on multiple-banded antigen (MBA) gene sequences.

Kong¹,

Ma²,

James³

et al. 2000

International Journal of Systematic and Evolutionary Microbiology

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Ureaplasma urealyticum has been divided into 14 serovars. Recently, subdivision of U. urealyticum into two species has been proposed : U. parvum (previously U. urealyticum parvo biovar), comprising four serovars (1, 3, 6, 14) and U. urealyticum (previously U. urealyticum T-960 biovar), 10 serovars (2, 4, 5, 7-13). The multiple-banded antigen (MBA) genes of these species contain both species and serovar/subtype specific sequences. Based on whole sequences of the 5'-ends of MBA genes of U. parvum serovars and partial sequences of the 5'-ends of MBA genes of U. urealyticum serovars, we previously divided each of these species into three MBA genotypes. To further elucidate the relationships between serovars, we sequenced the whole 5'-ends of MBA genes of all 10 U. urealyticum serovars and partial repetitive regions of these genes from all serovars of U. parvum and U. urealyticum. For the first time, all four serovars of U. parvum were clearly differentiated from each other. In addition, the 10 serovars of U. urealyticum were divided into five MBA genotypes, as follows : MBA genotype A comprises serovars 2, 5, 8 ; MBA genotype B, serovar 10 only ; MBA genotype C, serovars 4, 12, 13 ; MBA genotype D, serovar 9 only ; and MBA genotype E comprises serovars 7 and 11. There were no sequence differences between members within each MBA genotype. Further work is required to identify other genes or other regions of the MBA genes that may be used to differentiate U. urealyticum serovars within MBA genotypes A, C and E. A better understanding of the molecular basis of serotype differentiation will help to improve subtyping methods for use in studies of the pathogenesis and epidemiology of these organisms.

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Problems associated with serotyping strains of Ureaplasma urealyticum

Cited by 33 publications

References 20 publications

Simple Method for Determining Biovar and Serovar Types of Ureaplasma urealyticum Clinical Isolates Using PCR–Single-Strand Conformation Polymorphism Analysis

Simple Method for Determining Biovar and Serovar Types of Ureaplasma urealyticum Clinical Isolates Using PCR–Single-Strand Conformation Polymorphism Analysis

High-Resolution Melt PCR Analysis for Genotyping of Ureaplasma parvum Isolates Directly from Clinical Samples

Molecular genotyping of human Ureaplasma species based on multiple-banded antigen (MBA) gene sequences.

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