Clinical validation of a real-time polymerase chain reaction detection of Neisseria gonorrheae porA pseudogene versus culture techniques

Background: Diagnosing Neisseria gonorrheae using nucleic acid amplification tests (NAATs) might increase the sensitivity, compared to cultivation. However, using NAATs has also been problematic mainly due to the close genetic relationships between different Neisseria species, resulting in false positive diagnoses. This study was conducted to clinically validate a previously published real-time polymerase chain reaction (PCR) method targeting the porA pseudogene in N. gonorrheae in comparison to culture techniques.Methods: In total, 360 samples, urethra (n = 109), rectum (n = 84), pharynx (n = 119), and cervix (n = 48) from 185 males and 57 females, were analyzed using porA pseudogene PCR and cultivation. Sequencing of the entire porA pseudogene and the 16S rRNA gene were used to resolve discrepant results.Results: Of the 360 samples, 37 were positive by both culture and PCR, however, the PCR identified 15 additional confirmed positive samples. The PCR method showed a sensitivity, specificity, positive predictive value, and negative predictive value of 100% in a preselected population. The preselected population had a true gonorrhea prevalence of 17.4%.Conclusions: The present porA pseudogene real-time PCR comprises a valuable supplement to the traditional culture techniques for diagnosis of N. gonorrheae, especially for samples from extragenital sites such as pharynx and rectum.

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Mikrobiologi inom det medicinska området hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Microbiology in the medical area hsv//eng
Culture Techniques/*methods
DNA; Bacterial/analysis
Female
Gonorrhea/*diagnosis/microbiology
Humans
Male
Neisseria gonorrhoeae/*genetics
Polymerase Chain Reaction/*methods
Porins/*genetics
Predictive Value of Tests
Pseudogenes/genetics
MEDICINE
MEDICIN
Microbiology
Mikrobiologi
Microbiology
Mikrobiologi

In:Sexually Transmitted DiseasesPhiladelphia : American Venereal Disease Association35:5, s. 517-5200148-57171537-4521

By the author/editor: Hjelmevoll, Stig ...; Olsen, Merethe E ...; Sollid, Johanna ...; Haaheim, Håkon; Melby, Kjetil K.; Moi, Harald; show more...; Unemo, Magnus; Skogen, Vegard; show less...

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Basic Medicine; and Microbiology in ...

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