Evaluation of VIASURE Sexually Transmitted Diseases Real Time PCR Detection kit (CerTest Biotec) for the diagnostic of sexually transmitted infections

AbstractObjectivesTo develop a simple DNA sequencing test for simultaneous identification and antimicrobial resistance (AMR) detection of multiple sexually transmitted infections (STIs).MethodsA total of 200 vulvo-vaginal swab samples from 200 female sex workers in Ecuador were initially tested by real-time PCR for Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), Mycoplasma genitalium (MG) and Trichomonas vaginalis (TV). Samples positive for these STIs and controls were subjected to single gene targeted PCR MinION-nanopore sequencing by using the smartphone operated MinIT.ResultsAmong 200 vulvo-vaginal swab samples 43 were positive for at least one of the STIs by real-time PCR. Single gene targeted nanopore sequencing generally yielded higher pathogen specific reads in positive samples than negative controls. Of the 26 CT, NG or MG infections identified by real-time PCR, 25 were clearly distinguishable from the negative controls using sequence read count. Discrimination of TV positives from controls was poorer as many had low pathogen loads (cycle threshold > 35) with associated low sequence read counts. AMR analysis workflow indicated that 11% of the classified reads aligned with antibiotic resistance genes, all of which identified fluoroquinolone resistance in NG.ConclusionsSingle gene targeted nanopore sequencing for diagnosing and simultaneously identifying key antimicrobial resistance markers for four common STIs, associated with genital tract discharge in women shows promise. Further work to optimise accuracy, reduce costs and improve speed may allow sustainable approaches for managing STIs and emerging AMR in resource poor and laboratory limited settings.

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Accuracy of Curable Sexually Transmitted Infections and Genital Mycoplasmas Screening by Multiplex Real-Time PCR Using a Self-Collected Veil among Adult Women in Sub-Saharan Africa

Infectious Diseases in Obstetrics and Gynecology ◽

10.1155/2019/8639510 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15

Author(s):

Zita Aleyo Nodjikouambaye ◽

Fabrice Compain ◽

Damtheou Sadjoli ◽

Ralph-Sydney Mboumba Bouassa ◽

Hélène Péré ◽

...

Keyword(s):

Sexually Transmitted Infections ◽

Real Time ◽

Real Time Pcr ◽

Sub Saharan Africa ◽

Molecular Testing ◽

Adult Women ◽

Sampling Device ◽

Sexually Transmitted ◽

Sub Saharan ◽

Genital Mycoplasmas

Background. Sexually transmitted infections (STIs) are highly prevalent in sub-Saharan Africa. Genital self-sampling may facilitate the screening of STIs in hard-to-reach remote populations far from large health care centers and may increase screening rates. The cross-sectional GYNAUTO-STI study was carried out to assess the performance of a novel genital veil (V-Veil-Up Gyn Collection Device, V-Veil-Up Pharma, Ltd., Nicosia, Cyprus) as a genital self-sampling device to collect genital secretions to diagnose STIs by molecular biology as compared to reference clinician-collected genital specimens, in adult African women. Methods. Adult women living in N’Djamena, the capital city of Chad, were recruited from the community and referred to the clinic for women’s sexual health “La Renaissance Plus”. A clinician obtained an endocervical specimen using flocked swab. Genital secretions were also obtained by self-collection using veil. Both clinician- and self-collected specimens were tested for common curable STIs (including Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, and Trichomonas vaginalis) and genital Mycoplasma spp. by multiplex real-time PCR (Allplex™ STI Essential Assay, Seegene, Seoul, South Korea). Test positivities for both collection methods were compared by assessing methods agreement, sensitivity, and specificity. Results. A total of 251 women (mean age, 35.1 years) were prospectively enrolled. Only seven (2.8%) women were found to be infected with at least one common STIs [C. trachomatis: 3 (1.2%), N. gonorrhoeae: 1 (0.4%), M. genitalium: 4 (1.6%) and T. vaginalis: 1 (0.4%)], while the prevalence of genital mycoplasmas was much higher (54.2%) with a predominance of Ureaplasma parvum (42.6%). Self-collection by veil was non-inferior to clinician-based collection for genital microorganisms DNA molecular testing, with “almost perfect” agreement between both methods, high sensitivity (97.0%; 95%CI: 92.5-99.2%), and specificity (88.0%; 95%CI: 80.7-93.3%). Remarkably, the mean total number of genital microorganisms detected per woman was 1.14-fold higher in self-collected specimens compared to that in clinician-collected specimens. Conclusions. Veil-based self-collection of female genital secretions constitutes a convenient tool to collect in gentle way cervicovaginal secretions for accurate molecular detection of genital bacteria. Such sampling procedure could be easily implemented in STIs clinics in sub-Saharan Africa.

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