Date of Award

2008

Document Type

Master Thesis

Degree Name

Masters of Science (Research)

Department

Biological Sciences

First Advisor

Dr. Brigid Lucey

Abstract

The incidence of N. gonorrhoeae infections in Ireland has been steadily increasing since 2003. Culture is currently the preferred method of diagnosis, however the sensitivity of culture can be low. In recent years there has been increased interest in non-culture techniques such as polymerase chain reaction (PCR). Many of the PCR assays available have been shown to cross-react with commensal organisms, producing false positive results. This study aimed to improve the detection of N. gonorrhoeae by designing a multiplex PCR assay that used two N. gonorrhoeae genes as targets, thereby providing dual detection and confirmation of a positive result. PCR primers were designed to detect two N. gonorrhoeae genes, namely porA and pgil. Primers for an internal control were also designed. The V. cholerae ompW gene was chosen as the target for the internal control. The DNA of 51 organisms including 33 N. gonorrhoeae isolates, seven N. meningitidis isolates, two group B Streptococcus, a group A Streptococcus, an Enterococcus spp, a Coagulase negative Staphlycoccus and a Klebsiella pneumoniae were tested using the multiplex PCR assay. All 33 N. gonorrhoeae isolates were successfully detected by the assay while none of the nongonococcal isolates were detected. The assay therefore showed a specificity and sensitivity of 100%. The limit of detection was 5ng of DNA for this assay. This multiplex PCR assay therefore offers a sensitive and specific assay suitable for the detection of N. gonorrhoeae, and offers real potential for diagnostic use.

Access Level

info:eu-repo/semantics/openAccess

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