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A novel polymerase chain reaction assay to detect Mycoplasma genitalium.


ABSTRACT: AIMS:To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium. METHODS:Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine specimens from men with urethritis. RESULTS:The expected 341 bp product was produced on amplification of material from all M genitalium strains and from none of the other microorganisms tested. The lower limit of detection was 50 genome copies. The new assay detected M genitalium DNA in nine of 54 men with urethritis, in comparison with eight positive specimens detected with the alternative PCR. CONCLUSIONS:This novel PCR targeting the M genitalium 16S rRNA gene has been optimised and now provides a sensitive and specific alternative or addition to the available MgPa gene targeting assays.

SUBMITTER: Eastick K 

PROVIDER: S-EPMC1187285 | biostudies-literature | 2003 Feb

REPOSITORIES: biostudies-literature

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A novel polymerase chain reaction assay to detect Mycoplasma genitalium.

Eastick K K   Leeming J P JP   Caul E O EO   Horner P J PJ   Millar M R MR  

Molecular pathology : MP 20030201 1


<h4>Aims</h4>To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium.<h4>Methods</h4>Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine spec  ...[more]

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