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Eur J Clin Microbiol Infect Dis ; 33(1): 49-54, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23903665

RESUMO

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™ Q(x) 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™ Q(x) Collection Kit for Endocervical or Lesion Specimens.


Assuntos
DNA Bacteriano/isolamento & purificação , Gonorreia/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Neisseria gonorrhoeae/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Manejo de Espécimes/métodos , Proteínas da Membrana Bacteriana Externa/genética , DNA Bacteriano/genética , Humanos , Neisseria gonorrhoeae/genética , Porinas/genética , Sensibilidade e Especificidade
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