Erschienen in:
01.01.2014 | Article
Purification of DNA eluates from the ProbeTec GC Qx assay on BD Viper™ XTR allows for further analysis and confirmation of gonorrhea
verfasst von:
H. S. Tunsjø, E. Smedsrud, P. C. Holm, K. Rokvam, C. Schie, V. Rognlien, I. Augustin, A. Fostervold
Erschienen in:
European Journal of Clinical Microbiology & Infectious Diseases
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Ausgabe 1/2014
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Abstract
Low positive predictive values of Neisseria gonorrhoeae nucleic acid amplification tests in low-prevalence populations are a major challenge for accurate diagnostics. It is therefore necessary to verify all positive N. gonorrhoeae results with a different assay and target gene, preferably using the same sample. The BD ProbeTec™ Qx Collection Kit for Endocervical or Lesion Specimens, which is recommended for BD Viper™ XTR, is incompatible with other commercial platforms. Therefore, a confirmatory PCR has not been available for samples received on this transport medium. To be able to verify results from these samples with another assay, our objective was to establish a procedure for using the DNA eluates from BD Viper™ XTR for further analysis. DNA eluates from BD Viper™ XTR were collected and analyzed in two in-house confirmatory real-time PCRs targeting the porA pseudogene and the opa multicopy gene. BD Viper™ XTR DNA eluates were analyzed directly and also after purification with the nucleic acid extraction system NucliSENS® easyMag®. Purification of BD Viper™ XTR DNA eluates with the nucleic acid extraction system NucliSENS® easyMag® provided a sensitivity of the in-house PCR comparable to BD Viper™. With the inclusion of two target genes in the confirmatory PCR, specific and reliable verification of results were obtained. This study presents a simple, inexpensive procedure which allows for rapid verification of gonorrhea from samples received on the BD ProbeTec™ Qx Collection Kit for Endocervical or Lesion Specimens.