Retrospective Study for the Clinical Evaluation of a Real-Time PCR Assay with Lyophilized and Ready-to-Use Reagents for Streptococcus agalactiae Detection in Prenatal Screening Specimens.

Streptococcus agalactiae is a leading cause of sepsis and meningitis in newborns and young infants. Screening programs and intrapartum antibiotic prophylaxis have reduced early neonatal onset of disease. The aim of this study was to evaluate a molecular assay with lyophilized and ready-to-use reagents: VIASURE® Streptococcus B Real Time PCR detection kit (CerTest Biotec) (Viasure qPCR assay) compared to both the GBS culture and a molecular assay with separated and frozen reagents: Strep B Real-TM Quant (Sacace Biotecnologies®) (Sacace qPCR assay). A total of 413 vaginal−rectal swabs from women between the 35th and 37th weeks of pregnancy were processed. GBS culture was firstly achieved through Granada medium and Columbia CNA agar at 35 °C in aerobic conditions. Then, nucleic acid extraction was performed for subsequent molecular analysis using both commercial assays. Discordant results were resolved via bidirectional Sanger sequencing. Viasure qPCR assay clinical sensitivity was 0.97 (0.92−0.99) and specificity 1 (0.98−1). This retrospective study demonstrated the good clinical parameters and the strong overall agreement (99.3%) between the Viasure qPCR assay and both reference assays. Finally, the added value observed of the assay under study was the stabilized and ready-to-use format, reducing the number of time-consuming steps, permitting the storage at room temperature, facilitating transport, being environmentally respectful, and reducing additional costs.

Mycoplasma genitalium and sexually transmitted infections: evidences and figures in a tertiary hospital / Mycoplasma genitalium en el renovado diagnóstico de las infecciones de transmisión sexual: evidencias y cifras en un hospitalterciario

Introduction. Mycoplasma genitalium is an emerging cause of sexually transmitted infections (STIs) and has been implicated in non-gonococcal urethritis in men and cervicitis in woman. The aim of this study is determinate the incidence and pathogenicity of M. genitalium within the diagnosis of STIs detected from clinical samples in a third level hospital. Material and methods. A total of 8,473 samples from endocervix, urethra, vagina, rectum and others were processed applying Allpex STI Essential Assay. More than 190 records were reviewed to determinate M. genitalium pathogenicity. Results. M. genitalium was detected in a rate 2.8%. Co-infections were detected in 20% of the patients. Conclusions. M. genitalium is considered a STI emerging pathogen thanks to the renewal of multiplex-PCR tests although with a low incidence in our approach. Emerging from our experience and the institutional recommendations both detection of acid nucleic techniques (NAATs) and gonococcal culture might be implemented accurately and coexist to adequate prescriptions. (AU)

Introducción. Mycoplasma genitalium es un patógeno emergente causante de infecciones de transmisión sexual (ITS) y se ha relacionado con uretritis no gonocócica en hombres y cervicitis en mujeres. El objetivo de este estudio es determinar la incidencia y patogenicidad de M. genitalium en el seno del diagnóstico de ITS detectadas a partir de muestras clínicas en un hospital terciario. Métodos. Se procesaron 8.473 muestras de endocérvix, uretra, vagina, recto y otros, aplicando Allpex STI Essential Assay. Se revisaron más de 190 historias clínicas para determinar la patogenicidad de M. genitalium. Resultados. Se detectó M. genitalium en 2,8% de casos. Hubo coinfecciones en 20% de los pacientes. Conclusiones. M. genitalium a pesar de la baja incidencia en nuestra revisión, actualmente es un patógeno de valor en alza gracias al desarrollo de técnicas moleculares como PCRmultiplex. A partir de nuestra experiencia y las recomendaciones institucionales, tanto las técnicas de detección de ácidos nucleicos (NAATs) como los cultivos para gonococo deberían implementarse y coexistir para adecuar los tratamientos (AU)

Neisseria gonorrhoeae antimicrobial resistance in Spain: a prospective multicentre study.

OBJECTIVES: Gonococcal infection is one of the most reported sexually transmitted infections and antimicrobial resistance in Neisseria gonorrhoeae (NG) is challenging for the treatment of this infection. This observational study aimed to describe antimicrobial resistance of NG and epidemiological data from patients with gonococcal infection in eight regions of Spain, for updating the local therapeutic guidelines. METHODS: MICs of penicillin, cefixime, ceftriaxone, azithromycin, ciprofloxacin, fosfomycin and gentamicin were determined by Etest for all NG isolates recovered from 1 April 2018 to 30 September 2019 from 10 hospitals in Spain. Resistance determinants were identified using logistic regression analysis. Differences with a P value <0.05 were considered statistically significant. RESULTS: Antimicrobial susceptibility testing was performed for 2571 gonococci isolated from 2429 patients. 44.5% (945/2124) of patients were MSM. The resistance rate to extended-spectrum cephalosporins was low, with 0.2% (6/2561) of isolates resistant to ceftriaxone and 1.7% (44/2517) of isolates resistant to cefixime. The overall azithromycin resistance rate was 12.1% (310/2560), but differed greatly depending on the area. 56.2% (1366/2429) of the strains studied were ciprofloxacin resistant. MIC50 and MIC90 values of gentamicin and fosfomycin were 4 and 8 mg/L and 24 and 48 mg/L, respectively. CONCLUSIONS: Our study shows that NG susceptibility to extended-spectrum cephalosporins remains high in Spain. The azithromycin resistance rate questions the suitability of dual therapy. This study provides data of interest for updating the national treatment guidelines and highlights the need to develop and implement a national sentinel gonococcal antimicrobial susceptibility programme.

Update on vaginal infections: Aerobic vaginitis and other vaginal abnormalities / Actualización en infecciones vaginales: vaginitis aeróbica y otras alteraciones vaginales

It is estimated that abnormal vaginal discharge cannot be attributed to a clear infectious etiology in 15% to 50% of cases. Some women develop chronic vulvovaginal problems that are difficult to diagnose and treat, even by specialists. These disorders (aerobic vaginitis, desquamative inflammatory vaginitis, atrophic vaginitis, and cytolytic vaginosis) pose real challenges for clinical diagnosis and treatment. Researchers have established a diagnostic score based on phase-contrast microscopy. We review reported evidence on these entities and present our diagnostic experience based on the correlation with Gram stain. We recommend treatment with an antibiotic that has a very low minimum inhibitory concentration against lactobacilli and is effective against enterobacteria and Gram-positive cocci, which are responsible for these entities (aerobic vaginitis and desquamative inflammatory vaginitis)

Se estima que entre el 15 y el 50% de las mujeres que tienen trastornos del flujo vaginal, éstos no pueden atribuirse a una etiología infecciosa clara. Algunas de ellas desarrollarán problemas vulvovaginales crónicos difíciles de diagnosticar y tratar, incluso por especialistas. Son trastornos que plantean desafíos reales en el diagnóstico clínico y en su tratamiento como la vaginitis aeróbica, la vaginitis inflamatoria descamativa, la vaginitis atrófica y la vaginitis citolítica. Para diagnosticarlos, algunos investigadores han establecido una puntuación basada en la observación microscópica mediante contraste de fases. En este artículo, se revisa la evidencia publicada sobre estas entidades y presentamos nuestra experiencia en la correlación diagnóstica con la tinción de Gram. Se recomienda el tratamiento con un antibiótico con una concentración mínima inhibitoria muy baja contra los lactobacilos y eficaz contra las enterobacterias y los cocos grampositivos, responsables de estas entidades (vaginitis aeróbica y vaginitis inflamatoria descamativa)

High prevalence of spa types associated with the clonal lineage CC398 among tetracycline-resistant methicillin-resistant Staphylococcus aureus strains in a Spanish hospital.

OBJECTIVES: The clonal lineages, resistance mechanisms and virulence traits of tetracycline-resistant (Tet(R)) methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in a Spanish hospital during 2009 and 2010 were investigated. METHODS: Fifty-two Tet(R) MRSA strains from unrelated patients were included in this study. Susceptibility to 26 antimicrobial agents was determined and 24 resistance genes were tested for by PCR. The sequences of the genes grlA and gyrA were analysed in all ciprofloxacin-resistant MRSA isolates. For all strains, spa, SCCmec and agr typing was implemented. Multilocus sequence typing was performed for 16 representative strains of the different spa types. The presence of the genes tst, lukF/lukS-PV, eta, etb, etd and cna was investigated by PCR. RESULTS: Fifteen different spa types, four of them new ones, were detected among the 52 strains, being associated with the following clonal complexes (CCs): CC398 (67.3%), CC1 (11.5%), CC5 (11.5%) and CC8 (9.6%). A novel sequence type (ST), ST2077, belonging to CC398 was identified. Most MRSA CC398 strains were typed as SCCmecV-agrI. In addition to ß-lactam resistance, isolates showed resistance to (gene/number of strains): tetracycline [tet(K)/36, tet(L)/8 and tet(M)/48], macrolides and lincosamides [erm(B)/6, erm(C)/25, erm(T)/2, msr(A)/msr(B)/4 and mph(C)/4], aminoglycosides [aac(6')-Ie-aph(2')-Ia/8, ant(4')-Ia/13 and aph(3')-IIIa/15], trimethoprim [dfrS1/2 and dfrK/3] and mupirocin (mupA/3). Strains investigated for mutations mediating quinolone resistance revealed an S80F exchange in GrlA and different changes in GyrA. Three strains were Panton-Valentine leucocidin-positive (ST8 and ST94) and 41 strains were cna-positive. All MRSA isolates were negative for the genes tst, eta, etb and etd. CONCLUSIONS: Tetracycline resistance could be a good marker for MRSA CC398, although this resistance can also be found in other lineages.